Extended Protein Ions Are Formed by the Chain Ejection Model in Chemical Supercharging Electrospray Ionization.
Supercharging electrospray ionization can be a powerful tool for increasing charge states in mass spectra and generating unfolded ion structures, yet key details of its mechanism remain unclear. The structures of highly extended protein ions and the mechanism of supercharging were investigated using ion mobility-mass spectrometry. Head-to-tail-linked polyubiquitins (Ubq1-11) were used to determine size and charge state scaling laws for unfolded protein ions formed by supercharging while eliminating amino acid composition as a potential confounding factor. Collisional cross section was found to scale linearly with mass for these ions and several other monomeric proteins, and the maximum observed charge state for each analyte scales with mass in agreement with an analytical charge state scaling law for protein ions with highly extended structures that is supported by experimental gas-phase basicities. These results indicate that these highly unfolded ions can be considered quasi-one-dimensional, and collisional cross sections modeled with the Trajectory Method in Collidoscope show that these ions are significantly more extended than linear α-helices but less extended than straight chains. The effect of internal disulfide bonds on the extent of supercharging was probed using bovine serum albumin, β-lactoglobulin, and lysozyme, each of which contains multiple internal disulfide bonds. Reduction of the disulfide bonds led to a marked increase in charge state upon supercharging without significantly altering folding in solution. This evidence supports a supercharging mechanism in which these proteins unfold before or during evaporation of the electrospray droplet and ionization occurs by the Chain Ejection Model.


Functional characterization of grass carp runt-related transcription factor 3: Involvement in TGF-β1-mediated c-Myc transcription in fish cells.
In mammals, both runt-related transcription factor 3 (RUNX3) and c-Myc are the downstream effectors of transforming growth factor-β1 (TGF-β1) signaling to mediate various cellular responses. However, information of their interaction especially in fish is lacking. In the present study, grass carp (Ctenopharyngodon idella) runx3 (gcrunx3) cDNA was cloned and identified. Interestingly, opposing effects of recombinant grass carp TGF-β1 (rgcTGF-β1) on c-myc and runx3 mRNA expression were observed in grass carp periphery blood lymphocytes (PBLs). Parallelly, Runx3 protein levels were enhanced by rgcTGF-β1 in the cells. These findings prompted us to examine whether Runx3 can mediate the inhibition of TGF-β1 on c-myc expression in fish cells. In line with this, overexpression of grass carp Runx3 and Runx3 DN (a dominant-negative form of Runx3) in grass carp kidney cell line (CIK) cells decreased and increased c-myc transcript levels, respectively. Particularly, the regulation of Runx3 and Runx3 DN on c-myc mRNA expression was direct since they were presented in the nucleus without any stimulation. In addition, rgcTGF-β1 alone suppressed c-myc mRNA expression in CIK cells as in PBLs. Moreover, this inhibitory effect was also observed when grass carp Runx3 and Runx3 DN were overexpressed. These results strengthened the role of TGF-β1 signaling in controlling c-myc transcription. Taken together, TGF-β1-mediated c-myc expression was affected at least in part by Runx3, thereby firstly exploring the functional role of Runx3 in TGF-β1 down-regulation on c-myc mRNA expression in fish.


Enhanced hydrolysis of lignocellulosic biomass: Bi-functional enzyme complexes expressed in Pichia pastoris improve bioethanol production from Miscanthus sinensis.
Lignocellulosic biomass is the most abundant utilizable natural resource. In the process of bioethanol production from lignocellulosic biomass, an efficient hydrolysis of cellulose and hemicellulose to release hexose and pentose is essential. We have developed a strain of Pichia pastoris that can produce ethanol via pentose and hexose using an assembly of enzyme complexes. The use of enzyme complexes is one of the strategies for effective lignocellulosic biomass hydrolysis. Xylanase XynB from Clostridium cellulovorans and a chimeric endoglucanase cCelE from Clostridium thermocellum were selected as enzyme subunits, and were bound to a recombinant scaffolding protein mini-CbpA from C. cellulovorans to assemble the enzyme complexes. These complexes efficiently degraded xylan and carboxymethylcellulose (CMC), producing approximately 1.18 and 1.07 g/L ethanol from each substrate, respectively, which is 2.3-fold and 2.7-fold higher than that of the free-enzyme expressing strain. Miscanthus sinensis was investigated as the lignocellulosic biomass for producing bioethanol, and 1.08 g/L ethanol was produced using our recombinant P. pastoris strain, which is approximately 1.9-fold higher than that of the wild-type strain. In future research, construction of enzyme complexes containing various hydrolysis enzymes could be used to develop biocatalysts that can completely degrade lignocellulosic biomass into valuable products such as biofuels.


Interdisciplinary trauma management in an elderly patient, a case report.
The current report reviews a case of mixed dental trauma consequent to a fall by an older patient. The patient's teeth were forced out of alignment by the trauma and suffered pulpal necrosis. Treatment involved not only healing the acute injuries, but also attending to some subtle delayed problems that became apparent during treatment. Treatments involving endodontics, periodontics, orthodontics, and restorative dentistry were used to address all of the patient's concerns. This insured that the traumatic occlusion was corrected, appropriate esthetics was restored and normal speech and function was regained. All signs of trauma were recognized, every treatment step was documented, and appropriate follow-up was provided throughout the recovery period.


Civil society organisations and public health research--evidence from eight European union new member states.
Civil society organisations (CSO) are not-for-profit, non-governmental organisations operating in the public interest. They are the "third sector" that is strongly developed in Western European countries, ensuring the link between citizens and government and working as a counterbalance to the business sector. Their role in support of public health research deserves attention. Within a broader European study (STEPS--Strengthening Engagement in Public Health Research), public health organisations in eight European Union new member states (Cyprus, Estonia, Latvia, Lithuania, Malta, Romania, Slovakia, Slovenia) identified national CSOs with interests in health. A questionnaire was developed jointly, translated into national languages and sent by e-mail to 474 organisations, with 128 completed responses (27%). Most CSOs would like to be more involved in setting or advising on public-health research policy, and greater collaboration between CSOs, professional organisations and governmental institutions. Respondents did not see CSOs directly doing research, but recommended mobilizing researchers and organsations, supporting research themes, and lobbying to use public health evidence in policy and decision-making. They could receive more education for, and discussion of, public health research, and offer support in applying for research funding. Civil society organisations can contribute importantly,in setting public health research agendas. Research commissioning should give greater recognition of this role, improve links between CSOs, researchers and governmental institutions, and develop a stronger shared basis for public health policy and practice.


Comparative Investigation of Phenomenological Modeling for Hysteresis Responses of Magnetorheological Elastomer Devices.
Magnetorheological elastomer (MRE) is a type of magnetic soft material consisting of ferromagnetic particles embedded in a polymeric matrix. MRE-based devices have characteristics of adjustable stiffness and damping properties, and highly nonlinear and hysteretic force-displacement responses that are dependent on external excitations and applied magnetic fields. To effectively implement the devices in mitigating the hazard vibrations of structures, numerically traceable and computationally efficient models should be firstly developed to accurately present the unique behaviors of MREs, including the typical Payne effect and strain stiffening of rubbers etc. In this study, the up-to-date phenomenological models for describing hysteresis response of MRE devices are experimentally investigated. A prototype of MRE isolator is dynamically tested using a shaking table in the laboratory, and the tests are conducted based on displacement control using harmonic inputs with various loading frequencies, amplitudes and applied current levels. Then, the test results are used to identify the parameters of different phenomenological models for model performance evaluation. The procedure of model identification can be considered as solving a global minimization optimization problem, in which the fitness function is the root mean square error between the experimental data and the model prediction. The genetic algorithm (GA) is employed to solve the optimization problem for optimal model parameters due to its advantages of easy coding and fast convergence. Finally, several evaluation indices are adopted to compare the performances of different models, and the result shows that the improved LuGre friction model outperforms other models and has optimal accuracy in predicting the hysteresis response of the MRE device.


Comparative analysis of two C-terminal kinesin motor proteins: KIFC1 and KIFC5A.
We have taken advantage of the close structural relationship between two C-terminal motors, KIFC5A and KIFC1, to examine the sequence requirements for targeting of these two motors within the cell. Although KIFC5A and KIFC1 are almost identical in their motor and stalk domains, they differ in well-defined regions of their tail domains. Specific antisera to these motors were used to determine their localization to distinct subcellular compartments, the spindle for KIFC5A or membranous organelles for KIFC1. In addition to defining the intracellular localization of KIFC1, the reactivity of the KIFC1 antibody demonstrates that this motor contains a frame shift with respect to KIFC5A and is likely the product of a separate gene. The divergent tail domains of these motors are predicted to harbor specific information that directs them to their correct intracellular targets. In order to define the sequences responsible for the differential localization of these two motors, GFP was fused to motors with various tail deletions and their localization visualized after transfection. We were able to identify distinct sequences in each motor responsible for its unique cellular localization. The KIFC5A tail contains a 43 amino acid sequence with both nuclear localization and microtubule binding activity while KIFC1 contains a 19 amino acid sequence sufficient to target this motor to membrane-bounded organelles.


Gender differences in patients treated with the zotarolimus-eluting stent at a tertiary medical center.
Gender-related differences in long-term outcomes of patients receiving the Endeavor zotarolimus-eluting stent (ZES) (Medtronic) have not been well defined. In this study, we evaluate the differences between men (M) and women (W) for 2-year target vessel failure (TVF) in an unselected consecutive series of patients treated with the ZES at our institution. Data on 197 consecutive patients (133 M, 64 W) stented with the ZES were retrospectively analyzed. The primary endpoint of the study was to compare gender-related outcomes in TVF, defined as the combined endpoint of cardiac death, non-fatal myocardial infarction, and target vessel revascularization (TVR). Secondary endpoints included TLR, TVR, acute stent thrombosis (ST) as defined by the academic research consortium (ARC), and cardiac death. The cine angiograms of the first consecutive 122 patients (79 M, 43 W) were independently reviewed by a cardiologist blinded to clinical outcome and SYNTAX scoring was performed. Follow-up was achieved using medical records and/or phone calls and was censored at 730 days. Descriptive analysis was performed on all variables. Univariate analysis compared the M and W cohorts. Logistic regression analysis modeling for predictors of TVF was performed and survival analysis between the 2 groups was plotted. The 2 groups were well matched for demographic, clinical, angiographic, and procedural variables. Angiographic complexity was also statistically similar between the 2 groups as judged by SYNTAX scoring (15.8 ± 10.9 M vs 13.5 ± 8.3 W; P=.197). At 2-year follow-up, TVF was 22.6% and 32.8% (P=.684) with no statistical difference between TLR (18.1% M vs 12.8% W), TVR (21.8% M vs 32.8% W), cardiac death (2.3% M vs 6.3% W), and definite and probable stent thrombosis (2.26% M vs 3.13% W). Logistic regression analyses modeling for age, gender, New York Heart Association (NYHA) class, non-left main (LM) bifurcation lesions, ostial lesions, trifurcating LM, and pre-percutaneous coronary intervention (PCI) lesion severity showed that a higher NYHA class (odds ratio [OR], 2.68; P=.005), ostial lesions (OR, 5.68; P<.001), bifurcating non-LM lesions (OR, 2.74; P=.015), and trifurcating LM lesions (OR, 28.24; P<.001) predicted a higher TVF. Female gender (P=.086) and age (P=.09) were not independent predictors of TVF. In this cohort of patients receiving ZES, men and women had similar outcomes at 2-year follow-up consistent with recent reports in the current era of PCI. Complex coronary anatomy (ostial, non-LM bifurcations, and LM trifurcations) and advanced heart failure were stronger predictors of higher TVF than gender and age.


Progesterone receptor isoform A may regulate the effects of neoadjuvant aglepristone in canine mammary carcinoma.
Progesterone receptors play a key role in the development of canine mammary tumours, and recent research has focussed on their possible value as therapeutic targets using antiprogestins. Cloning and sequencing of the progesterone receptor gene has shown that the receptor has two isoforms, A and B, transcribed from a single gene. Experimental studies in human breast cancer suggest that the differential expression of progesterone receptor isoforms has implications for hormone therapy responsiveness. This study examined the effects of the antiprogestin aglepristone on cell proliferation and mRNA expression of progesterone receptor isoforms A and B in mammary carcinomas in dogs treated with 20 mg/Kg of aglepristone (n = 22) or vehicle (n = 5) twice before surgery. Formalin-fixed, paraffin-embedded tissue samples taken before and after treatment were used to analyse total progesterone receptor and both isoforms by RT-qPCR and Ki67 antigen labelling. Both total progesterone receptor and isoform A mRNA expression levels decreased after treatment with aglepristone. Furthermore, a significant decrease in the proliferation index (percentage of Ki67-labelled cells) was observed in progesterone-receptor positive and isoform-A positive tumours in aglepristone-treated dogs. These findings suggest that the antiproliferative effects of aglepristone in canine mammary carcinomas are mediated by progesterone receptor isoform A.


[Evaluation of therapeutic efficacy by detection of specific IgG and IgG4 against KLH in schistosomiasis japonica].
The specific IgG and IgG4 antibodies in the sera of patients with Schistosoma japonicum were determined by KLH-ELISA and SEA-ELISA. The results showed that KLH-ELISA was as sensitive and specific as SEA-ELISA. The negative conversion rates of IgG and IgG4 12 months after treatment detected by KLH-ELISA were significantly higher than those by SEA-ELISA. The negative conversion rate of IgG was 92.9%, and that of IgG4 was 97.6% 24 months after treatment. Therefore, it is regarded that detecting specific IgG and IgG4 by KLH has high value in both diagnosing schistosomiasis japonica and assessing its therapeutic efficacy. Particularly the specific IgG4 may be a short-duration antibody which can indicate successful chemotherapy of patients with chronic Schistosoma japonicum infection.


Comparison of the WISC-R and culture fair intelligence test in a juvenile delinquent population.
Culture Fair and WISC-R scores were obtained of 51 delinquent juveniles. Comparison of mean scores on the two tests for minority and white juveniles demonstrated the Culture Fair to be less culturally biased than the WISC-R. On analysis of variance there was a significant difference due to ethnicity and test used, and a significant interaction of those two factors. The interaction was due to the elevation of the score of minority juveniles on the Culture Fair compared with WISC-R scores. Correlations between the WISC-R scores, subtest scale scores, and Culture Fair scores were significant at the .01 level which provides further validation of the WISC-R.


Effect of temperature on growth and reproduction of the epigeic earthworm, Eudrilus Eugeniae (Kinberg).
Influence of temperature on growth and reproduction of Eudrilus eugeniae has been investigated by laboratory culturing at regulated 25.0 degrees C, 30.0 degrees C, 37.5 degrees C and 40.0 degrees C and in fluctuating (22.7-27.3 degrees C) room temperature of prevailing (winter) season over 16 weeks. All worms died during first and tenth week at 40.0 degrees C and 37.5 degrees C respectively. Weight (biomass) and growth of worms cultured at different temperature varied significantly (P<0.01). The mean growth (mg/g live weight/day) at 25.0 degrees C., 30.0 degrees C, 35.0 degrees C, 37.5 degrees C and in fluctuating temperatures was 1,074.04 +/- 6.07, 1,554.01 degrees 192.37, 148.1 +/- 15.28, 192.83 +/- 25.8 and 1450.4 +/- 162.1 respectively. Growth declined after maturity drastically with coccon production. At 25.0 degrees C though worms are sexually mature, they failed to produce cocoons within 16 weeks whereas, at 35.0 degrees C and 37.5 degrees C they did not sexually mature. Worms attained sexual maturity at a mean weight of about 1000 mg/worm. The mean per cent maturity was higher and earlier in fluctuating temperatures and at 30.0 degrees C than at 25.0 degrees C. Cocoon production was observed only at 30.0 degrees C and in fluctuating temperatures with a mean of 0.9 and 1.5 cocoons/ wom/week and the cumulative cocoon number of 10.8 and 14.7/worm over 16th week respectively. The fluctuating temperature of uncontrolled room environment and 30.0 degrees C were favorable for various life activities of the worms. Eudrilus eugeniae appears to have range of temperature optima more than 25.0 degrees C and less than 28 degrees C. The climatic conditions prevailing in whole of the peninsular India during winter season are favourable for employing this worm in intensive field scale vermiculturing practices.


Starch Modification with Molecular Transformation, Physicochemical Characteristics, and Industrial Usability: A State-of-the-Art Review.
Starch is a readily available and abundant source of biological raw materials and is widely used in the food, medical, and textile industries. However, native starch with insufficient functionality limits its utilization in the above applications; therefore, it is modified through various physical, chemical, enzymatic, genetic and multiple modifications. This review summarized the relationship between structural changes and functional properties of starch subjected to different modified methods, including hydrothermal treatment, microwave, pre-gelatinization, ball milling, ultrasonication, radiation, high hydrostatic pressure, supercritical CO2, oxidation, etherification, esterification, acid hydrolysis, enzymatic modification, genetic modification, and their combined modifications. A better understanding of these features has the potential to lead to starch-based products with targeted structures and optimized properties for specific applications.


99mTc-LHRH in tumor receptor imaging.
Detection of gonadotropin-releasing hormone (GnRH) also known as luteinizing hormone-releasing hormone (LHRH) in the relevant tumor tissue and normal tissues and organs in vivo expression was investigated. To examine the method of direct radio labeling of LHRH by 99mTc with relatively high radiochemical purity and stability, screening the best labeling conditions, to establish a simple and reliable method of preparation of 99mTc-LHRH was undertaken. The detection of radioisotope-labeled LHRH distribution in mice, LHRH receptor imaging for the study and treatment of cancer basis were evaluated. i) Immunohistochemical staining test was used in 23 patients with hepatocellular carcinoma (HCC), 20 patients with breast cancer, 10 patients with prostate cancer, 20 patients with lung cancer, 20 patients with endometrial cancer tumor cells and normal tissue LHRH-R De Biaoda levels; ii) pre-tin method use direct labeling of LHRH, marking completion of saline or human serum were added at room temperature, the chromatography was measured at different times, to calculate the rate of labeled product and the radiochemical purity of the label, in vivo observation of its stability, and comparative analysis of selected optimal condition; iii) rat pituitary cell membrane protein, the product of in vitro radio-receptor marker analysis, through the saturation and inhibition experiments, was used to test its receptor binding activity; iv) Ch-T method labeled 125I-LHRH, tail vein injection of normal mice at different times were sacrificed, blood and major organs were determined and calculated per gram organization percentage injected dose rate (%, ID/g). Detected by immunohistochemistry in 23 cases of HCC in the LHRH-positive rate was 82.61%, in the corresponding normal tissues, the positive rate was 15%; 20 cases of breast cancer positive rate of 95%, the corresponding normal tissues, the positive rate was 20%; 10 cases of prostate cancer positive rate of 70%, the corresponding normal tissues, the positive rate of 40%; 20 cases of lung cancer positive rate of 85%, the corresponding normal tissues, the positive rate of 15.79%; 20 cases of endometrial cancer positive rate of 80% in the corresponding normal tissues was 16.67% positive. 99mTc-LHRH mark was 97.9-100.0%, the radiochemical purity of 93.9-96.4%, marking the reaction gel content of <5%. Great product receptor marker analysis showed 99mTc-LHRH with saturable receptor binding characteristics and inhibition, and high affinity, RT = 23.2174 pmol, KD = 0.4348 nmol; intravenous injection of 131I-LHRH within 72 h after the mice rapidly cleared the blood radioactivity, the major radioactive accumulation in the liver and kidneys and by the liver, renal clearance, and other tissues and organs of the radioactivity gradually decreased with time. In conclusion, i) the liver, lung, breast, prostate, endometrial cancer exist in both LHRHR; ii) 99mTc-LHRH preparation is simple, rapid, radiochemical purity product obtained higher marks, better stability, no further purification; and iii) LHRH 99mTc labeled, still has a high receptor binding ability, biological activity; and has an ideal and realistic dynamics in animals, there is hope, as with the clinical value of imaging agent of GnRH receptors.


Small subunit ribosomal DNA phylogeny of microsporidia that infect Daphnia (Crustacea: Cladocera).
Glugoides intestinalis, Microsporidium sp., Ordospora colligata, Gurleya vavrai, Larssonia obtusa and Flabelliforma magnivora are microsporidian parasites of planctonic freshwater crustaceans Daphnia spp. We performed a phylogenetic analysis of the small subunit ribosomal DNA which revealed their positions as polyphyletic. G. intestinalis, Microsporidium sp. and O. colligata, which are horizontally transmitted gut parasites with small spores and low virulence, group with different lineages. G. intestinalis is related to 2 microsporidia infecting lepidopterans and to Vittaforma corneae, which has been described as a human pathogen. It is thought that V. corneae may have an invertebrate as its natural host. Microsporidium sp. is a relative of the genera Enterocytozoon and Nucleospora, pathogens of man and fish respectively. O. colligata is the first species found to be closely related to the genus Encephalitozoon, which is comprised of 3 species that are parasites of homeothermic vertebrates. G. vavrai and L. obtusa are sister taxa that branch close to the Amblyosporidae, the only microsporidia with known intermediate hosts. This finding supports the presumption of secondary hosts for G. vavrai and L. obtusa, as it has not been possible to maintain these species in Daphnia in the laboratory. F. magnivora roots deep at the base of the phylum microsporidia with no close relative found so far.


Evaluation of a new fiber-reinforced resin composite.
Efficacy of the usage of an experimental fiber-reinforced composite (FRC) on mechanical properties of an indirect composite was investigated by means of three-point bending and Charpy impact tests. Bond strength between the FRC and the indirect composite was also evaluated by tensile testing. The FRC consisted of a matrix resin with 25% silanized milled glass fiber (11-microm diameter, 150-microm length) and 5% colloidal silica. The values of strain of proportional limit, total strain, and fracture energy of the FRC during the bending test (1.2%, 10.4%, and 41.6 x 10(-3) J) were significantly higher than those of the indirect composite (0.1%, 2.5%, and 11.9 x 10(-3) J). The impact strengths of the 1-mm specimens with FRC ranged from 15.2 to 15.9 kJ/m(2), and were significantly higher than that of the control (3.1 kJ/m(2)). The 2-mm specimens showed significant difference from the control when the FRC thickness was equal or greater than 0.5 mm. The bond strength after the thermocycling was 15.2 MPa, and all of the specimens exhibited cohesive fracture inside the indirect composite. Based upon the results, it was concluded that the FRC tested in this study improved toughness and impact resistance of the indirect composite. The interfacial bonding between the FRC and the indirect composite was strong enough to prevent delamination.


Staph wars: the antibiotic pipeline strikes back.
Antibiotic chemotherapy is widely regarded as one of the most significant medical advancements in history. However, the continued misuse of antibiotics has contributed to the rapid rise of antimicrobial resistance (AMR) globally. Staphylococcus aureus, a major human pathogen, has become synonymous with multidrug resistance and is a leading antimicrobial-resistant pathogen causing significant morbidity and mortality worldwide. This review focuses on (1) the targets of current anti-staphylococcal antibiotics and the specific mechanisms that confirm resistance; (2) an in-depth analysis of recently licensed antibiotics approved for the treatment of S. aureus infections; and (3) an examination of the pre-clinical pipeline of anti-staphylococcal compounds. In addition, we examine the molecular mechanism of action of novel antimicrobials and derivatives of existing classes of antibiotics, collate data on the emergence of resistance to new compounds and provide an overview of key data from clinical trials evaluating anti-staphylococcal compounds. We present several successful cases in the development of alternative forms of existing antibiotics that have activity against multidrug-resistant S. aureus. Pre-clinical antimicrobials show promise, but more focus and funding are required to develop novel classes of compounds that can curtail the spread of and sustainably control antimicrobial-resistant S. aureus infections.


FOXP3 induced by CD28/B7 interaction regulates CD25 and anergic phenotype in human CD4+CD25- T lymphocytes.
Among the signals necessary to generate CD4(+)CD25(+)FOXP3(+) T cells from CD4(+)CD25(-)FOXP3(-) T cells, a pivotal role is played by CD28. However, in humans, it is not known whether CD28 signaling independently of TCR promotes forkhead box protein 3 (FOXP3) expression and regulates CD4(+)CD25(+)FOXP3(+) T cell functions. To address this issue, starting from our previous experience, we analyzed the unique signals delivered by CD28 following stimulation by its natural ligand B7. Our results show that, in primary CD4(+)CD25(-) T cells, CD28 signals independent of TCR-mediated stimulatory pathways are sufficient to induce the transcription of FOXP3 in a small number of CD4(+)CD25(-) T cells committed to express FOXP3. These signals are dependent on CD28-derived PI3K/Akt pathways and resistant to cyclosporin A. In addition, we demonstrated that translated FOXP3 was recruited to CD25, Il-2, and Ctla4 target promoters. CD28-mediated FOXP3 expression was transient and correlated with CD25 expression. The presence of FOXP3 in CD28-activated CD4(+)CD25(-) T cells correlated with a transient unresponsiveness to antigenic stimuli. The addition of exogenous IL-2 did not influence either FOXP3 or CD25 expression but rescued CD28-activated T cells from apoptosis. Our results, demonstrating that FOXP3 expression driven solely by the CD28/B7 interaction inhibited T cell activation, support the role of CD28 in the regulation of peripheral tolerance and suggest a new mechanism through which it could occur.


Effect of arsenic trioxide on the Tregs ratio and the levels of IFN-γ, IL-4, IL-17 and TGF-β1 in the peripheral blood of severe aplastic anemia patients.
Previous studies have suggested that the anticancer agent, arsenic trioxide (ATO), could attenuate T cell mediated immunity by not only inhibiting the proliferative response of T cells but by also increasing the frequency of regulatory T cells (Tregs). Furthermore, ATO represents a reasonable salvage treatment in some patients with refractory severe aplastic anemia (SAA). The current study aimed to evaluate the function of ATO on the Tregs percentage and cytokines changes in the peripheral blood mononuclear cells (PBMCs) of SAA patients.PBMCs were collected from 20 newly diagnosed SAA patients in Henan Cancer Hospital and treated with different concentrations of ATO (0, 1, 2.5, and 5 μmol/L). Then we investigated the efficacy of ATO on Tregs ratio and the levels of interferon (IFN)-γ, interleukin (IL)-4, IL-17 and transforming growth factor (TGF)-β1 in the peripheral blood of SAA patients in vitro.The results showed that ATO significantly increased the proportion of Tregs (P < .001) at 2.5 and 5 μmol/L concentrations, and the proportion of Tregs was increased with increasing ATO concentration (r = 0.524). At 1 (P = .03), 2.5 (P < .001) and 5 μmol/L (P < .001), ATO significantly up-regulated the expression levels of Foxp3 mRNA, which was positively and linearly correlated with the increase of Tregs cell-frequency (r = 0.52, 95%CI, 0.37-0.67). In addition, ATO significantly reduced the levels of IFN-γ (at 1, 2.5 and 5 μmol/L, P < .001), IL-4 (at 2.5 μmol/L, P = .009; at 5 μmol/L, P < .001), and IL-17 (at 2.5, P = .016; at 5 μmol/L, P < .001). ATO significantly reduced the levels of TGF-β1 at 5 μmol/L (P = .03), but showed no significant effects at 1 and 2.5 μmol/L (P > .05).ATO could mediate the immune regulation, which might contribute to improve hematopoietic recovery in SAA patients.


The development of quality indicators for home care in China.
To develop a comprehensive system of quality indicators for home care in China. A modified Delphi technique and analytic hierarchy process. Twenty experts were invited to participate in the Delphi expert consultation. Experts rated the perceived importance of 92 potential indicators through two rounds of e-mail surveys in November and December 2016. The analytic hierarchy process was used to determine the relative importance of the quality indicators identified through the Delphi expert consultation. The average authoritative coefficient was 0.815 ± 0.0432 (range: 0.75-0.90). After two rounds of Delphi expert consultation, 77 quality indicators were identified as important in the Chinese home care setting. The mean importance ratings ranged from 4.35 to 4.95 on a 5-point scale, with variation coefficients ranging from 0.04 to 0.15. The percentage of experts giving the maximum possible score on each item ranged from 45% to 95%. In the second round, the Kendall's W coefficients ranged from 0.363 to 0.570. As for relative importance, the weights were 0.198-0.490, 0.029-0.047 and 0.037-0.66 for first-level, secondary-level and third-level quality indicators, respectively, with all consistency ratio values less than 0.1. The absolute and relative importance of 77 indicators identified as potentially valid measures of the quality of Chinese home care was determined. This instrument is the first set of home care quality indicators developed specifically for mainland China, and it should be useful in evaluating and improving the quality of Chinese home care.


The management of paediatric neurogenic bladder: an approach in a resource-poor setting.
If untreated, paediatric neurogenic bladder can cause renal failure and urinary incontinence. It is usually caused by neural tube defects such as myelomeningocele. Children with a neurogenic bladder should be monitored from birth and management should aim to preserve renal function and achieve social continence. This article outlines the management options appropriate for these children in resource-poor settings. In most low- and middle-income countries, a general lack of awareness of the neurological effects on the urinary tract results in late presentation, usually with urological complications even when spina bifida is diagnosed early. Physical examination must include neurological examination for spinal deformities and intact sacral reflexes. About 90% of children with occult spinal dysraphisms will have cutaneous sacral lesions. The work-up includes urinalysis, serial ultrasound of the urinary tracts and urodynamics. Urodynamic assessment is essential for the diagnosis and prognosis of the paediatric neurogenic bladder. In poorly resourced settings, simple eyeball urodynamics can be performed in the absence of a conventional urodynamic set-up. Clean intermittent catheterisation (CIC), the mainstay of treatment, is most suitable for resource-poor settings because it is effective and inexpensive. Antimuscarinic drugs such as oxybutynin complement CIC by reducing detrusor overactivity. Intravesical injection of Botox and bladder augmentation surgery is required by a small subset of patients who fail to respond to combined CIC and oxybutynin therapy. Children with neurogenic bladder in resource-poor settings should have early bladder management to preserve renal function and provide social continence.


Urea transporter knockout mice and their renal phenotypes.
Urea transporter gene knockout mice have been created for the study of the urine-concentrating mechanism. The major findings in studies of the renal phenotype of these mice are as follows: (1) Urea accumulation in the inner medullary interstitium is dependent on intrarenal urea recycling mediated by urea transporters; (2) urea transporters are essential for preventing urea-induced osmotic diuresis and thus for water conservation; (3) NaCl concentration in the inner medullary interstitium is not significantly affected by the absence of IMCD, descending limb of Henle and descending vasa recta urea transporters. Studies in urea transporter knockout mouse models have highlighted the essential role of urea for producing maximally concentrated urine.


Evaluation of the antibacterial activity of cystatin against selected strains of Escherichia coli.
The aim of this study was to analyze the antibacterial activity of hen egg white cystatin against selected Escherichia coli strains. We used a monomeric solution of hen egg white cystatin in bovine serum albumin (BSA) with added phosphate buffered saline (PBS), and three test strains: Escherichia coli ATCC 23811, Escherichia coli ATCC 8739 and Escherichia coli ATCC 25922. The effect of cystatin against the tested strains was determined on the basis of minimal inhibitory concentration (MIC) and survival curves of the microorganisms in a cystatin-containing environment during incubation at various temperatures. Our study confirmed the activity of cystatin against the analyzed Escherichia coli strains. taining environment, as compared to control samples incubated in a ovocystatin-deficient medium. Depending on the incubation temperature (20 degrees C or 37 degrees C) the reduction persisted up to 12 hours after incubation.


Long-range gamma phase synchronization as a compensatory strategy during working memory in high-performing patients with schizophrenia.
Working memory deficits in schizophrenia may be associated with impairments in the integration of neural activity across a distributed network of cortical areas. However, evaluation of the contribution of this integration to working memory impairments in patients is severely confounded by behavioral performance. In the present multidimensional-neuroimaging study, measures of neural oscillations at baseline and during a working memory task, baseline gamma-aminobutyric acid (GABA) level in the left dorsolateral prefrontal cortex (DLPFC), and behavioral performance were obtained. Controlling behavioral performance by recruiting only "high-performing" patients with schizophrenia, we investigated whether the strength of cross-area communications differs between patients with schizophrenia and healthy participants under accurate and equivalent behavioral performance. Results of phase-locking value indicated that these high-performing patients recruited significantly more between frontal and occipital regions in the left hemisphere, t(13) = -2.16, p = .05, Cohen's d = -1.20, and between frontal and temporal regions in the right hemisphere, t(13) = -2.63, p = .02, Cohen's d = -1.46. These cross-area communication patterns may be associated with visuoverbal and visuospatial working memory networks of the left and right hemispheres, respectively. Moreover, correlations of patient's cross-area communication with in vivo GABA levels of the left DLPFC revealed a significant positive relationship (r = .77, p = .04), demonstrating that the critical role of GABA functions in gamma band oscillations may go beyond local neuronal assemblies in the left DLPFC. Altogether, these exploratory findings point to the heterogeneity among schizophrenia patients and highlight the notion that high-performing patients may engage in potential compensatory mechanisms and may represent a subgroup of patients that may be categorically or dimensionally divergent in psychopathology.


Comparing oral lithium carbonate and intraperitoneal lithium chloride chronic administrations on rats' activity levels.
This study compares the effects produced by two modalities of lithium administration on rats' activity levels. Lithium was administered for 21 days either as carbonate in the diet (2 g in 2 liters of water and 1,500 g lab chow) or as chloride through single daily intraperitoneal injections (1 mEq/kg LiCl). Both treatments resulted in similar decreases in spontaneous activity and exploratory behavior, as recorded in an open field with a hole-board. Neither treatment influenced the amount of food consumed on a daily basis nor the rate of growth as manifested by weight gains over the period considered, suggesting that these treatments were not adversely affecting the animals' health. Both treatments increased water intake over control levels, this effect being most marked with the diet lithium carbonate administration, particularly during the second week of treatment. This latter modality of lithium administration, but not the injections of lithium chloride, significantly increased NaCl intake over control levels.


The lived experience of Lebanese family caregivers of cancer patients.
The purpose of this study was to explore, through in-depth semistructured interviews, the lived experience of Lebanese family caregivers of cancer patients and acquire a better knowledge of the meaning and interpretation of their experience. The study design was based on the Utrecht School of Phenomenology. This study followed purposeful sampling, in which 9 participants with a mean age of 51 years were selected. Data were analyzed using the hermeneutic phenomenological approach based on the Utrecht School of Phenomenology. Eight core themes describing the participants' lived experience emerged from the interviews: living with fears and uncertainty, loss of happiness, feeling of added responsibility, living in a state of emergency, sharing the pain, living the dilemma of truth telling, disturbed by being pitied, and reliance on God. The results of this study challenge nurses to be conscious of the nature and difficulties that family caregivers are encountering.


[The absence of monocyte-granulocyte antigen expression in a case of AML].
The brief record of a 25 y. o. male patient with AML (FAB M1) is shown, in whom the blast cells did not express any of the 5 myeloid antigens or the other-lineage related antigens, as detected with the monoclonal antibodies. The blast cells were induced to express CD13 antigen after a short-term culture in vitro. This result suggests that CD13 antigen can be expressed virtually by all AML cells, since CD13 antigen is known to cover fresh AML cells at the highest incidence. No unusual clinical feature was noted in this patient as AML case. The collected documentation of antigen-free AML cases seems necessary for the relevant understanding of AML heterogeneity.


Prognostic Value of Serum Biomarkers of Autoimmunity for Recurrence of IgA Nephropathy after Kidney Transplantation.
A prognostic biomarker for IgA nephropathy (IgAN) recurrence after renal transplant is lacking. We followed 96 consecutive first renal transplant recipients with native kidney IgAN (79 men; 92 deceased donors; mean age =48.1 years) on calcineurin inhibitor-based immunosuppression over 10 years for death, allograft failure, and clinicopathologic recurrence (CPR; clinically evident and biopsy-proven). Using time-dependent Cox regression analysis and receiver operating characteristic curves, we assessed prognostic significance of levels of galactose-deficient IgA1 (Gd-IgA1; autoantigen) and Gd-IgA1-specific IgG and IgA autoantibodies in serum obtained at time of transplant or native-kidney IgAN diagnosis (30 patients only). Overall, 13 patients died, 34 kidneys failed (17 due to CPR), and 34 patients developed CPR after a mean interval of 5.8 years. Compared with healthy controls (n=30), patients had significantly elevated serum Gd-IgA1 levels at diagnosis and transplant, but levels did not associate with any outcome. Patients also had significantly elevated levels of normalized (but not total) serum Gd-IgA1-specific IgG autoantibodies at diagnosis and transplant, and the level at transplant associated with higher risk of CPR (relative risk, 2.68; 95% confidence interval, 1.26 to 5.71; P=0.01; area under the receiver operating characteristic curve, 0.62; 95% confidence interval, 0.51 to 0.74; P=0.05). Normalized Gd-IgA1-specific IgG autoantibody level remained an independent risk factor for CPR in multivariate analysis. Serum Gd-IgA1-specific IgA autoantibody level did not change between diagnosis and transplant or predict outcome. This study emphasizes post-transplant prognostic value of normalized serum IgG antiglycan autoantibody level in patients with IgAN.


Enantiomeric separation of 13 new amphetamine-like designer drugs by capillary electrophoresis, using modified-B-cyclodextrins.
An easy-to-prepare chiral CE method for the enantiomeric separation of 13 new amphetamine-like designer drugs, using CDs as chiral selectors, was developed. Sulfated-β-CD was found to be the best chiral selector among the three used (sulfated-β-CD, caroboxymethyl-β-CD, dimethyl-β-CD). The separation of the analytes was achieved in a fused-silica gel capillary at 20 °C using an applied voltage of +25 kV. The optimized background electrolyte consisted of 63.5 mM H3 PO4 and 46.9 mM NaOH in water. Several electrophoretic parameters such as CD type, CD concentration (1 - 40 mg/mL), buffer pH (2.6, 3.6, 5.0, 6.0), length of the capillary (70 - 40 cm total length), amount of the organic solvent (methanol and acetonitrile) were investigated and optimized.


[Protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against oxidative injury].
The protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against ox-LDL-induced damage to human umbilical vein endothelial cells (HUVECs) was investigated by MTT assay, and the components were identified by using UPLC-Q-TOF-MS. According to the study, ethyl acetate extract and n-butanol extract could increase cell viability (P<0.01), while petroleum ether extract had no influence, and water extract could even inhibit the cell viability to some degree. Moreover, 32 compounds in four polar fractions were analyzed, including 31 quinones and their glycosides, and one rubiprasins C. Petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract contained 23, 32, 26, 15 compounds, respectively. According to cell experiments in vitro, active fractions were ethyl acetate extract and n-butanol extract. The results could provide scientific references for further studies on effective material basic of cRRR, and lay a foundation for studies on the relationship between efficacies and materials.


Correlation matrix of a completely polarized, statistically stationary electromagnetic field.
It is shown that, for a 3 x 3 correlation matrix Wij(r, r, omega), (i, j = x, y, z) of the electric vector of a random, stationary electromagnetic field to represent a field that is completely polarized at a point r and frequency omega, each element of the matrix must factorize. More precisely, a necessary and sufficient condition for the correlation matrix to represent a fully polarized field at a point r is that the matrix has the form Wij(r, r, omega) = epsilon(i)*(r, omega)epsilon(j)(r, omega), where epsilon(i)(r, omega) (i = x, y, z) are deterministic functions, i.e., that all pairs of the Cartesian components of the electric field at a point r and frequency omega are completely correlated.


Against medical ethics: opening the can of worms.
In a controversial paper, David Seedhouse argues that medical ethics is not and cannot be a distinct discipline with it own field of study. He derives this claim from a characterization of ethics, which he states but does not defend. He claims further that the project of medical ethics as it exists and of moral philosophy do not overlap. I show that Seedhouse's views on ethics have wide implications which he does not declare, and in the light of this argue that Seedhouse owes us a defence of his characterization of ethics. Further, I show that his characterization of ethics, which he uses to attack medical ethics, is a committed position within moral philosophy. As a consequence of this, it does not allow the relation between moral philosophy and medical ethics to be discussed without prejudice to its outcome. Finally, I explore the relation between Seedhouse's position and naturalism, and its implications for medical epistemology. I argue that this shows us that Seedhouse's position, if it can be defended, is likely to lead to a fruitful and important line of inquiry which reconnects philosophy and medical ethics.


New challenges and opportunities in child welfare outcomes and information technologies.
In spite of longstanding concern on the part of child welfare policymakers, researchers, and practitioners, we still lack basic information about the outcomes of child welfare services. This article describes the limitations of current efforts to generate data on child welfare outcomes, presents certain principles for the development of child welfare management information systems for the 21st century, and suggests steps to be taken to get from here to there. Particular attention is given to recent federal initiatives that provide an immediate opportunity to make a fundamental leap in our ability to understand the outcomes of child welfare services.


Highlights of the 22nd French pharmacovigilance meeting.
(1) The 22nd French pharmacovigilance meeting, held in July 2001, presented data on adverse effects notified by health professionals to regional pharmacovigilance centres in France. (2) Non specific "immunostimulants" are not harmless placebos, as might be concluded from the relative lack of data. There have been 315 notifications of severe adverse effects, some with positive rechallenge, reporting cutaneous, gastrointestinal, respiratory, haematological and other disorders. Attributability was considered "likely" in 68% of cases. Three deaths occurred. (3) Other well known adverse effects continue to occur: convulsions with camphor, visual hallucinations with oxybutynin, headache with antimigraine drugs, liver damage with dextropropoxyphene, neuropsychological disorders after buflomedil overdose (especially in patients with renal failure), hyperkalaemia during spironolactone combination with an angiotensin-converting-enzyme inhibitor (ACE inhibitor), and severe infections after intravesical BCG. (4) Rare adverse effects of old drugs were identified, such as oedema with valproic acid, interstitial pneumonia with flecainide, and a bleeding risk due to tramadol interaction with oral anticoagulants. (5) The adverse effects of new drugs are better documented: celecoxib is now implicated in visual disorders. (6) Overall, the meeting confirmed that only a small proportion of adverse drug reactions are notified, that a large number of hospitalised patients suffer from drug induced complications, and that summaries of product characteristics (SPC) are often too brief or reassuring regarding pharmacovigilance data. The poor risk-benefit ratios of some drugs call for their immediate market withdrawal.


Calcium-mediated C(sp3)-H Activation and Alkylation of Alkylpyridines.
Main-group metal calcium-mediated alkylpyridine benzylic C(sp3)-H activation and functionalization have been achieved. The reaction of a calcium hydride complex [{(DIPPnacnac)CaH(thf)}2] (DIPPnacnac = CH{(CMe)(2,6-iPr2-C6H3N)}2) with two equivalents of 2,6-lutidine rapidly yields a monomeric calcium alkyl complex with the release of dihydrogen. A hydride/carbon-bridged binuclear calcium complex [{(DIPPnacnac)Ca}2(μ-H){2-Me-6-(μ-CH2)-Py}(thf)] is obtained from an equimolar treatment of calcium hydride and 2,6-lutidine that is readily converted into mono- or binuclear calcium alkyl complexes upon subsequent addition of 2,6-lutidine. DFT calculations and kinetic studies are conducted to determine their reaction profiles. More significantly, this calcium hydride complex catalyzes regioselective benzylic C-H bond addition of alkylpyridines to a variety of alkenes, affording linear or branched alkylated pyridine derivatives.


Novel Cancer Stem Marker and Its Applicability for Grading Primary Human Gliomas.
Poorly differentiated cell populations including tumor-initiating stem cells have been demonstrated to display a unique ability to natively internalize fragmented double-stranded DNA. Using this feature as a marker, we show that 0.1% to 6% of human glioblastoma cells from the bioptates can effectively internalize a fluorescently labeled DNA probe. Of these, using samples from 3 patients, 66% to 100% cells are also positive for CD133, a well-established surface marker of tumor-initiating glioma stem cells. Using the samples from primary malignant brain lesions (33 patients), we demonstrate that tumor grading significantly correlates ( R = .71) with the percentage of DNA-internalizing cells. No such correlation is observed for relapse samples (18 patients).


Aerobic C-H Oxidation of Arenes Using a Recyclable, Heterogeneous Rhodium Catalyst.
A novel, practical protocol for the aerobic direct C-H acetoxylation of arenes, employing a recyclable heterogeneous rhodium catalyst, is reported herein. The trifluoroacetoxylation of 2-amido-substituted anthracenes proceeded at the 9-position with exclusive regioselectivity. The oxidation of variously substituted anthracenes and other polycyclic aromatics with molecular oxygen as a terminal oxidant proceeded under mild conditions, providing products in good to excellent yields.


Effect of urokinase antiserum on plasminogen activators: demonstration of immunologic dissimilarity between plasma plasminogen activator and urokinase.
Antiserum against purified human urokinase was produced by immunization of Hartley strain guinea pigs. The antiserum was capable of neutralizing the plasminogen activator activity of the antigen and of native urokinase in human urine. The antiserum did not inhibit plasminogen activators of bacterial origin, i.e., streptokinase and staphylokinase; neither did it inhibit urokinase from nonprimate mammals, i.e., dog, pig, rabbit, guinea pig, nor tissue activator or tissue culture supernatants from porcine sources. Partial cross-reactivity against urokinase from primates, i.e., rhesus monkey and baboon, was noted as well as with supernatant from rhesus kidney tissue culture. In vitro studies showed lack of immunologic identity between human urokinase and human milk activator or human tissue activator from adrenal sources but demonstrated immunologic identity between human urokinase and the supernatant from human kidney tissue culture. In vivo studies in man failed to show detectable levels of urokinase activity in peripheral venous or renal venous blood under a variety of clinical states and when stimuli such as exercise, electroshock therapy, or nicotinic acid were used to enhance plasminogen activator activity in the plasma. The results establish that human plasma activator, milk activator, and tissue activator from the adrenals are immunologically distinct from human urokinase.


Combinatorial protocol in multiple linear regression/partial least-squares directed rationale for the caspase-3 inhibition activity of isoquinoline-1,3,4-trione derivatives.
The caspase-3 inhibition activity of isoquinoline-1,3,4-trione derivatives has been analysed with the topological and molecular features from Dragon software. Analysis of the structural features in conjunction with the biological endpoints in combinatorial protocol in multiple linear regression (CP-MLR) led to the identification of 45 descriptors for modelling the activity. The study clearly suggested the role of rotatable bonds, mean information on the distance degree equality, radial centricity, bond and structural information content of five-order neighbourhood symmetry, atomic van der Waals volumes and the presence or absence of certain structural fragments to optimise the caspase-3 inhibitory activity of titled compounds. The models developed and the participating descriptors advocate that the substituent groups of the isoquinoline moiety hold scope for further modification in the optimization of the caspase-3 inhibitory activity. Analysis of these descriptors in partial least squares (PLS) highlighted their relative significance in modulating the biological response. The selected descriptors are enriched with information corresponding to the activity when compared to the remaining ones.


Prevalence, influencing factors, screening and diagnosis of delirium at stationary hospitalised patients in a Swiss acute Hospital. A pilot study
Background: Deliria have a massive effect on patients, from increased duration of hospitalization to higher mortality. Risk factors such as age, deprivation of substances, immobility as well as stress are known among others. Particularly in vulnerable persons minor factors can lead to a delirium. European studies report a prevalence rate between 17 % and 22 %, but can't be compared to the Swiss hospital system. No national delirium prevalence data in acute hospitals is known. Aim: On the one hand to measure the delirium prevalence in an acute hospital, to elaborate patient characteristics of delirium patients based on group comparison and to test sensitivity and specifity of the applied instruments, on the other hand to get information about the practicality of the study execution. Method: Delirium point prevalence measurement has been conducted in a prospective cross-sectional study. On one determined day data of patients have been collected by nurses in an acute hospital. Results: A prevalence point rate of 14 % (6 / 43) based on CAM has been identified. Significant differences were found between the groups in respect of age, discipline, number of ICD diagnoses, care dependency and in all the three delirium instruments. Delirium patients were not only longer hospitalized but had almost twice as many ICD diagnosed, were high-maintenance patients and mostly didn't claim to be in pain. Conclusion: This is the first prevalence study in a Swiss acute hospital. The utilized instruments are reliable and the study execution is practicable and could be conducted with a larger sample. Most known risk factors were confirmed.


Variability of immunologic and clinical features in patients with rheumatoid arthritis studied over 24 hours.
Disease activity measures in patients with rheumatoid arthritis (RA) are influenced by many factors. Bedrest has been long recognized to effect decreased disease activity. In addition, most patients note maximal levels of pain and stiffness in the early morning hours with improvement in the afternoon. Cytokines and hormones with circadian rhythms may also contribute to variability in disease status during the day. Our purpose was to investigate variability of clinical and immunologic measures over a 24-h period of hospitalization in patients with RA. Production of rheumatoid factors and immunoglobulins, joint counts and questionnaire evaluations were measured 3 to 6 times over a 24-h period of hospitalization in 10 patients with RA who were not receiving steroids or second line drugs. Total joint counts were significantly decreased as soon as 4 h after admission, and continued to decrease during the 24-h period of observation. Significant decreases in mononuclear cell proliferation and antibody synthesis were also observed. Our findings are consistent with the presence of decreased levels of immunostimulatory mediators in hospitalized patients especially during late afternoon or evening hours.


Beneficial effects of long-term use of the antioxidant probucol in heart failure in the rat.
Congestive heart failure (CHF) is a disease that is characterized by progressive left ventricular (LV) dysfunction and dilatation. Oxidative stress is thought to contribute to the progression of CHF, and antioxidants have been shown to have beneficial effects when started early after myocardial infarction (MI). In this study, we tested whether the powerful antioxidant probucol would attenuate progression of CHF once it was established after MI in the rat. Ligation of a coronary artery was used to create an MI in rats (n=266). Survivors were then randomized 20 days after MI to either probucol 61 mg. kg(-1). d(-1) or vehicle and followed up for a total of 100 days after MI. Studies of cardiac hemodynamics, LV remodeling, cardiac apoptosis and morphology, systemic neurohumoral activation, oxidative stress, and renal function were then evaluated. Probucol improved LV function (LV maximum rate of pressure rise from 3103 to 4250 mm Hg/s, P<0.05, and LV end-diastolic pressure decrease from 28 to 24 mm Hg, P<0.05), reduced pulmonary weights, prevented right ventricular systolic hypertension, and preserved renal function compared with vehicle. Probucol also prevented LV dilatation, prevented wall thinning (1.70 versus 1.42 mm, P<0.05), reduced cardiac fibrosis and cardiac apoptosis, attenuated increased myocardial cell cross-sectional area, and increased scar thickness. In chronic CHF, probucol exerts multiple beneficial morphological effects that result in better LV remodeling and function, reduced neurohumoral activation, and preserved renal function.


The implications of high bed occupancy rates on readmission rates in England: A longitudinal study.
Hospital bed occupancy rates in the English National Health Service have risen to levels considered clinically unsafe. This study assesses the association of increased bed occupancy with changes in the percentage of overnight patients discharged from hospital on a given day, and their subsequent 30-day readmission rate. Longitudinal panel data methods are used to analyse secondary care records (n = 4,193,590) for 136 non-specialist Trusts between April 2014 and February 2016. The average bed occupancy rate across the study period was 90.4%. A 1% increase in bed occupancy was associated with a 0.49% rise in the discharge rate, and a 0.011% increase in the 30-day readmission rate for discharged patients. These associations became more pronounced once bed occupancy exceeded 95%. When bed occupancy rates were high, hospitals discharged a greater proportion of their patients. Those were mostly younger and less clinically complex, suggesting that hospitals are successfully prioritising early discharge amongst least vulnerable patients. However, while increased bed occupancy was not associated with a substantial increase in overall 30-day readmission rates, the relationship was more pronounced in older and sicker patients, indicating possible links with short-fallings in discharge processes.


Scleral Proteome in Noninfectious Scleritis Unravels Upregulation of Filaggrin-2 and Signs of Neovascularization.
Scleritis is a severe inflammatory ocular disorder with unknown pathogenesis. We investigated healthy sclera as well as sclera affected by noninfectious scleritis for differentially expressed proteins using a mass spectrometry approach. We collected scleral samples of enucleated eyes due to severe noninfectious scleritis (n = 3), and control scleral tissues (n = 5), all exenterated eyes for eyelid carcinomas (n = 4), or choroidal melanoma (n = 1) without scleral invasion. Samples were prepared for the nano liquid-chromatography mass spectrometer (LC-MS), data were analyzed using proteomics software (Scaffold), and is available via ProteomeXchange (identifier PXD038727). Samples were also stained for immuno-histopathological evaluation. Mass spectrometry identified 629 proteins within the healthy and diseased scleral tissues, whereof collagen type XII, VI, and I were the most abundantly expressed protein. Collagen type II-XII was also present. Filaggrin-2, a protein that plays a crucial role in epidermal barrier function, was found upregulated in all scleritis cases. In addition, other epithelial associated proteins were upregulated (such as keratin 33b, 34, and 85, epiplakin, transglutaminase-3, galectin 7, and caspase-14) in scleritis. Further, upregulated proteins involved in regulation of the cytoskeleton (vinculin and myosin 9), and housekeeping proteins were found (elongation factor-2 and cytoplasmic dynein 1) in our study. Upregulation of filaggrin-2 and myosin-9 was confirmed with immunohistochemistry, the latter protein showing co-localization with the endothelial cell marker ETC-related gene (ERG), indicating neovascularization in scleral tissue affected by scleritis. We found upregulation of filaggrin-2 and signs of neovascularization in scleral tissue of patients with noninfectious scleritis. Further research, ideally including more scleritis cases, is needed to validate our findings.


Overcoming Obstacles in the Development of Antigen-Specific Immunotherapies for Type 1 Diabetes.
Antigen-specific immunotherapy (ASI) holds great promise for type 1 diabetes (T1D). Preclinical success for this approach has been demonstrated in vivo, however, clinical translation is still pending. Reasons explaining the slow progress to approve ASI are complex and span all stages of research and development, in both academic and industry environments. The basic four hurdles comprise a lack of translatability of pre-clinical research to human trials; an absence of robust prognostic and predictive biomarkers for therapeutic outcome; a need for a clear regulatory path addressing ASI modalities; and the limited acceptance to develop therapies intervening at the pre-symptomatic stages of disease. The core theme to address these challenges is collaboration-early, transparent, and engaged interactions between academic labs, pharmaceutical research and clinical development teams, advocacy groups, and regulatory agencies to drive a fundamental shift in how we think and treat T1D.


[Interleukin-10 in laryngeal cancer patients treated by surgery or radiotherapy].
The early detection and right monitoring of the laryngeal cancer make a better chance of recovery. Therefore factors that might be showing the presence of neoplastic process are constantly being looked for. The detection of correlation between incidence of cancer and immunity system disorders makes us look for and try to answer an important question: are there any factors that might be cancer markers in immunity system? Cytokines are molecules that regulate the process of differentiation and proliferation. They are also the mediators of immunological and inflammatory reactions. Interleukina-10 (IL-10) is a cytokine that has various important functions stimulating immunity processes. IL-10 is a factor stimulating killer cells/cytoxic lymphocytes/ and is able to kill cancer cells growth spontaneously. It has been detected that some cancer cells can produce some IL-10, too. The aim of the study was to evaluate the serum level concentration of IL-10 in the laryngeal cancer patients before any medical treatment and its denotation after surgery or radiotherapy. A group of 43 male patients with laryngeal cancer was considered for the purpose of this study. 22 patients were treated by surgery and 21 by radiotherapy. In all the cases the serum concentration of IL-10 before the treatment, in the first and 30th days after therapy was evaluated. The results were compared with the group of 20 healthy volunteers. The mean value of IL-10 concentration in laryngeal cancer patients treated by surgery was 14.0 pg/ml before treatment, 12.8 pg/ml in the 1 day and 14.4 pg/ml 30 days after the therapy. IL-10 concentrations in patients treated by radiotherapy were respectively: 15.4, 14.4 and 15.9 pg/ml. The mean value of IL-10 in the control group was 4.9 pg/ml. The serum concentrations of IL-10 in the laryngeal cancer patients were significantly higher than in the control group. These IL-10 concentrations didn't change significantly after the surgery or radiotherapy. The serum concentration of IL-10 in the laryngeal cancer patients is significantly higher than in the healthy control group. The changes of serum concentrations after the surgery or radiotherapy of laryngeal cancer patients are not significant and characteristic.


Wide local excision could be considered as the initial treatment of primary anorectal malignant melanoma.
Anorectal malignant melanoma was a rare disease with extremely poor prognosis. The aim of this study was to explore the clinical characteristic, diagnosis and treatment strategies of anorectal malignant melanoma. The data of 57 patients with anorectal malignant melanoma was collected and retrospectively analyzed. Rectal bleeding and anal mass were found to be common symptoms of anorectal malignant melanoma. The preoperative diagnosis rate of anorectal malignant melanoma was 48.6%. The overall 3-year and 5-year survival rate was 38.0% and 21.3% respectively. The 3-year survival rates of stage I and II patients were 63.0% and 16.7% respectively (P = 0.000), and the 5-year survival rates were 33.3% and 11.1% (P = 0.001), which both had significant statistic differences. The 3-year survival rate of patients undergone abdmoninoperineal resection and patients undergone wide local excision were 36.7% and 53.0% respectively (P = 0.280), while the 5-year survival rate were 24.1% and 23.1% (P = 0.642), which both had no significant statistic differences. This study identified no survival advantage to abdominoperineal resection in treatment of anorectal malignant melanoma, and we propose that wide local excision could be considered as the initial treatment of choice.


Increasing health-care options: the perspectives of parents who use complementary and alternative medicines.
To explore the relationship between conventional medicine and complementary and alternative medicine (CAM) with parents who use CAM, and to consider factors that may contribute to parent non-disclosure of CAM usage to their doctor. Thirty-three parents participated in one of seven focus groups. Transcripts were analysed using an iterative process of theme identification and testing against transcript data. The participants believed they should trust their instincts as parents in caring for their child. It was important also to the participants that they understood why their child was ill, and a range of theories of health and illness were discussed. The use of CAM was attractive as it offered more options in health care than just relying on conventional medicine alone. The use of additional therapies was seen as a means to increase the likelihood that something would work. Many of the participants described bad experiences with doctors when they discussed CAM use previously so they had become more circumspect in mentioning it. The participants were most satisfied with medical care for their child when they felt the doctor respected their point of view and listened to them. Doctors caring for children and their families should expect that many parents are using CAM to increase health-care options. Inquiries about CAM usage should be made in a non-judgmental and encouraging manner so parents feel comfortable in providing an honest answer. Advice to parents about CAM may need to be backed up with evidence to address differing parent understanding of illness.


Sick leave of spouses to cancer patients before and after diagnosis.
The impact of cancer on spouses of cancer patients may be considerable in many aspects. Our objective was to evaluate sick leave in spouses of cancer patients before and after the diagnosis. Using Swedish population-based registries, we studied sick leave of spouses to patients with newly diagnosed colon, rectal, lung, prostate, or breast cancer. We identified the cancer patients via the Swedish Cancer Registry and obtained information of their spouse through linkage with the population register. We assessed the number of sick leave episodes and sick days one year before until one year after the spouses' cancer diagnosis by cross-referencing with Swedish Social Insurance Agency data. We also compared the number of sick days of spouses with the general population adjusted for age, sex and partner status. In general, spouses (N=1 923) to cancer patients had an increase in the frequency of new episodes of sick leave in the months before and after the cancer diagnosis. Spouses of lung cancer patients had most sick leave episodes, and the largest number of sick days per person. In comparison to the general population, spouses in the lung cancer group also had the highest standardised sick day ratio 1.76; 95% confidence interval 1.24, 2.40. The corresponding risk for spouses in other groups of cancer was not significantly increased. In Sweden there is often increased sick leave of spouses to cancer patients. It may be due to emotional stress and physical reactions that follow with cancer which needs to be further explored in order to provide adequate support and care.


Use of the protein A-gold immunocytochemical and enzyme-gold cytochemical techniques in studies of vitellogenesis.
Vitellogenesis in the frog hepatocyte was investigated by applying the protein A-gold immunocytochemical and RNase-gold cytochemical techniques in conjunction with morphometric and biochemical analyses. The morphometric studies demonstrated that the surface density of rough endoplasmic reticulum (RER) and nucleolar size increased more than fourfold and 1.25-fold, respectively, while the nuclear size and the mitochondrial compartment size remained constant following estrogen treatment. Concurrently, liver RNA concentration increased 2.5-fold while protein and DNA concentrations did not change. In addition, total plasma protein more than doubled, with vitellogenin accounting for 40% of the final volume. The secretory proteins vitellogenin and protein-RcX (a nonvitellogenin, estrogen-induced plasma protein of unknown function, found in the plasma of Rana catesbeiana) were detected immunocytochemically in the RER, Golgi apparatus, and secretory granules in hepatocytes only of estrogen-treated frogs. Lysosomes also were labeled. These observations established that protein-RcX was synthesized and secreted by the hepatocyte in parallel with vitellogenin and that both of these export proteins were confined to the secretory pathway and lysosomes. Quantitation of labeling density indicated that the concentration of vitellogenin increased as it progressed along the secretory vector. Albumin was detected immunocytochemically also within these same hepatocyte entities from both untreated and treated animals. In the untreated animals, albumin concentration also increased progressively along the secretory vector. A marked alteration of albumin processing was observed following estrogen treatment. While albumin concentration in the RER was unchanged, its concentrations within the Golgi apparatus and secretory granules were lower than those observed in the RER or in counterpart compartments under control conditions. RNase-gold cytochemistry for total RNA demonstrated a 1.5-fold increase in labeling density over the nucleolus but no change in RER labeling following estrogen treatment. These labeling data, in combination with the morphometric data, suggest an increase of approximately 80% in the total amount of RNA in the nucleolus and 430% in the RER in response to estrogen. This review thus illustrates the significant contributions which can be made by gold-probe techniques, alone or in combination with morphometric and biochemical techniques, to investigations of the intracellular processing of secretory proteins.


Unregulated LDL cholesterol uptake is detrimental to breast cancer cells.
Tumor uptake of exogenous cholesterol has been associated with the proliferation of various cancers. Previously, we and others have shown that hypercholesterolemia promotes tumor growth and silencing of the LDL receptor (LDLR) in high LDLR-expressing tumors reduces growth. To advance understanding of how LDL uptake promotes tumor growth, LDLR expression was amplified in breast cancer cell lines with endogenously low LDLR expression. Murine (Mvt1) and human (MDA-MB-468) breast cancer cell lines were transduced to overexpress human LDLR (LDLROE). Successful transduction was confirmed by RNA and protein analysis. Fluorescence-labeled LDL uptake was increased in both Mvt1 and MDA-MD-468 LDLROE cells. The expression of the cholesterol-metabolizing genes, ABCA1 and ABCG1, was increased, while HMGCR was decreased in the MDA-MB-468 LDLROE cells. In contrast, Mvt1 LDLROE cells showed no differences in Abca1 and Abcg1 expression and increased Hmgcr expression. Using a Seahorse analyzer, Mvt1 LDLROE cells showed increased respiration (ATP-linked and maximal) relative to controls, while no statistically significant changes in respiration in MDA-MB-468 LDLROE cells were observed. Growth of LDLROE cells was reduced in culture and in hypercholesterolemic mice by two-fold. However, the expression of proliferation-associated markers (Ki67, PCNA and BrdU-label incorporation) was not decreased in the Mvt1 LDLROE tumors and cells. Caspase-3 cleavage, which is associated with apoptosis, was increased in both the Mvt1 and MDA-MB-468 LDLROE cells relative to controls, with the Mvt1 LDLROE cells also showing decreased phosphorylation of p44/42MAPK. Taken together, our work suggests that while additional LDL can promote tumor growth, unregulated and prolonged LDL uptake is detrimental.


Toxicity assessment and detoxification metabolism of sodium pentachlorophenol (PCP-Na) on marine economic species: a case study of Moerella iridescens and Exopalaemon carinicauda.
Sodium pentachlorophenol (PCP-Na) is widespread in the marine environment; however, its impact on marine organisms remains under-researched. Moerella iridescens and Exopalaemon carinicauda are marine species of economic importance in China and under threat from PCP-Na pollution. Thus, this study aimed to assess the toxicity and detoxification metabolism of PCP-Na on M. iridescens and E. carinicauda. The study revealed that the 96 h median lethal concentration (LC50) of PCP-Na for M. iridescens and E. carinicauda were 9.895 mg/L and 14.143 mg/L, respectively. A species sensitivity distribution (SSD) for PCP-Na was developed specifically for marine organisms, determining a hazardous concentration to 5% of the species (HC5) of 0.047 mg/L. During the sub-chronic exposure period, PCP-Na accumulated significantly in M. iridescens and E. carinicauda, with highest concentrations of 41.22 mg/kg in the soft tissues of M. iridescens, 42.58 mg/kg in the hepatopancreas of E. carinicauda, and only 0.85 mg/kg in the muscle of E. carinicauda. Furthermore, the study demonstrated that detoxifying metabolic enzymes and antioxidant defense system enzymes of E. carinicauda responded stronger to PCP-Na compared to M. iridescens, suggesting that E. carinicauda may possess a stronger detoxification capacity. Notably, five biomarkers were identified and proposed for monitoring and evaluating PCP-Na contamination. Overall, the results indicated that M. iridescens and E. carinicauda exhibit greater tolerance to PCP-Na than other marine species, but they are susceptible to accumulating PCP-Na in their tissues, posing a significant health risk. Consequently, conducting aquatic health risk assessments in areas with potential PCP-Na contamination is strongly recommended.


A pilot study to compare the use of prototypes of multipositional paediatric walking sticks and tripods with conventional sticks and tripods by children with cerebral palsy.
The researchers (B.H. and B.T.) designed prototypes of a walking stick and a tripod termed Multipositional Paediatric Walking Aids for children with cerebral palsy (CP). The design won the 'Jenx Award for Innovation in Paediatric Physiotherapy'. The object of this study was to ascertain whether the prototypes had any benefit over conventional designs in children with CP. A small case series within-subject comparison design was used. There were four periods, AABA, each 4 weeks in length. Prototypes were used during period B. Eight children with CP between 4 and 11 years were recruited, four used sticks and four used tripods. Following an orthopaedic assessment, each child had assessments at the commencement of the study and every 4 weeks thereafter: (1) Energy used while walking using Physiological Cost Index (PCI). (2) Motor abilities using (a) Gross Motor Function Measure (GMFM-88) and (b) Gross Motor Performance Measure (GMPM). (3) Hand/forearm position recorded on a visual analogue. (4) Parent/child questionnaire. Physiological Cost Index improved overall when the prototypes were used suggesting the amount of energy used when walking with the prototypes was less than with conventional sticks/tripods. GMFM-88 and GMPM results improved for some but not all children. Results for PCI, GMFM and GMPM were not statistically significant. The hand/forearm position of stick users consistently improved at assessment 4. Questionnaire results indicated that stick users preferred the prototypes. Multipositional sticks/tripods may have benefits over conventional sticks/tripods. Children using sticks rather than tripods preferred the prototypes. The study demonstrates the need to undertake scientifically controlled trials before marketing new equipment.


Erratum.
The first two sentences of the report "Asian dust: Seasonal transport to the Hawaiian Islands" by J. R. Parrington et al. (8 Apr., p. 195) should have read as follows: "Transport of vast quantities of Sahara desert dust westward across the Atlantic Ocean to Bermuda has been known since the late 1960's (1, 2). Strong evidence now exists for transport of large quantities of Asian dust eastward across the North Pacific (3-7)."


"This is where it all started" - the pivotal role of PLCζ within the sophisticated process of mammalian reproduction: a systemic review.
Mammalian reproduction is one of the most complex and fascinating biological phenomenon, which aims to transfer maternal and paternal genetic material to the next generation. At the end of oogenesis and spermatogenesis, both haploid gametes contain a single set of chromosomes ready to form the zygote, the first cell of the newly developing individual. The mature oocyte and spermatozoa remain in a quiescent state, during which the oocyte is characterized by nuclear and cytoplasmic arrest, while the spermatozoa necessitates further maturation within the epididymis and female reproductive track prior to egg fertilization. Either in vivo or in vitro, the sperm initiates a series of irreversible biochemical and physiological modifications in the oocyte. The earliest detected signal after fertilization is cytosolic Ca2+ oscillations, a prerequisite step for embryo development. These oscillations trigger the release of the oocyte from the second meiosis arrest towards embryogenesis, also known as "oocyte activation". Phospholipase C zeta (PLCζ) is a unique sperm-soluble protein responsible for triggering the InsP3/Ca2+ pathway within the oocyte, leading to Ca2+ oscillations and consequently to embryo development. The specific structure of PLCζ (compared to other PLCs) enables its specialized activity via the preserved X and Y catalytic domains, as well as distinct features such as rapid onset, high sensitivity to Ca2+ and cession of oscillations upon zygote formation. The emerging discoveries of PLCζ have stimulated studies focusing on the possible clinical applications of this protein in male infertility evaluation and management during IVF/ICSI. Fertilization failure is attributed to lack of oocyte second meiosis resumption, suggesting that ICSI failure may be related to impaired PLCζ activity. Microinjection of recombinant human PLCζ to human oocytes after ICSI fertilization failure may trigger Ca2+ oscillations and achieve successful fertilization, offering new hope for couples traditionally referred to sperm donation. However, more studies are still required prior to the routine implementation of this approach in the clinic. Directions for future studies are discussed.


Melatonin Attenuates Calcium Deposition from Vascular Smooth Muscle Cells by Activating Mitochondrial Fusion and Mitophagy via an AMPK/OPA1 Signaling Pathway.
Mitochondrial fusion/mitophagy plays a role in cardiovascular calcification. Melatonin has been shown to protect against cardiovascular disease. This study sought to explore whether melatonin attenuates vascular calcification by regulating mitochondrial fusion/mitophagy via the AMP-activated protein kinase/optic atrophy 1 (AMPK/OPA1) signaling pathway. The effects of melatonin on vascular calcification were investigated in vascular smooth muscle cells (VSMCs). Calcium deposits were visualized by Alizarin Red S staining, while calcium content and alkaline phosphatase (ALP) activity were used to evaluate osteogenic differentiation. Western blots were used to measure expression of runt-related transcription factor 2 (Runx2), mitofusin 2 (Mfn2), mito-light chain 3 (mito-LC3) II, and cleaved caspase 3. Melatonin markedly reduced calcium deposition and ALP activity. Runx2 and cleaved caspase 3 were downregulated in response to melatonin, whereas Mfn2 and mito-LC3II were enhanced and accompanied by decreased mitochondrial superoxide levels. Melatonin also maintained mitochondrial function and promoted mitochondrial fusion/mitophagy via the OPA1 pathway. However, OPA1 deletion abolished the protective effects of melatonin on VSMC calcification. Melatonin treatment significantly increased p-AMPK and OPA1 protein expression, whereas treatment with compound C ablated the observed benefits of melatonin treatment. Collectively, our results demonstrate that melatonin protects VSMCs against calcification by promoting mitochondrial fusion/mitophagy via the AMPK/OPA1 pathway.


Right brachial access is safe for branched endovascular aneurysm repair in complex aortic disease.
The risk of perioperative cerebrovascular events in endovascular repair of thoracic and thoracoabdominal aneurysms is reported from 2% to 15%. The unavoidable use of an upper extremity access during branched endovascular aneurysm repair (b-EVAR) may play a role in embolic brain injuries. For this reason, some advocate the use of a left-sided upper access to avoid crossing the origin of supra-aortic vessels. However, the assumption that right brachial access has a higher risk for stroke during b-EVAR has not been confirmed in the literature. This study retrospectively analyzed all consecutive patients treated by b-EVAR with right brachial access at a single institution. A through-and-through right-brachiofemoral 0.014-inch wire was used to stabilize the sheath across the arch in all cases. End point of the study was the incidence of cerebrovascular events. We identified 61 patients (65.6% male) during a 4-year period. Mean age at the time of surgery was 70.4 years (range, 53-87 years). The most common indication for treatment was type II (32.8%), followed by type IV thoracoabdominal aortic aneurysms (23%). There were 20 urgent (32.8%) and 41 elective (67.2%) procedures. Two perioperative ischemic strokes occurred in the first postoperative day in two men (3.3%; 95% confidence interval, 0.397-11.84). No further ischemic strokes occurred perioperatively. There was no statistically significant association between the occurrence of postoperative stroke and any of the perioperative characteristics. No significant association was found between the duration of the procedure and the end point. In both patients with embolic events, the use of a left arm approach would not have been feasible due to coverage of the left subclavian artery ostium. The postoperative stroke rate in b-EVAR with the use of a right brachial access in our experience was in line with the literature for treatment of thoracic and thoracoabdominal aortic aneurysms. We conclude that the right brachial access with the use of a stabilizing through-and-through wire is a safe approach during b-EVAR.


Responses to continuous and intermittent exercise in healthy and insulin-dependent diabetic children.
The hormonal (growth hormone, glucagon, cortisol) and metabolic (glucose, ketone bodies) responses to 30 min of continuous vs 30 min of intermittent exercise were evaluated in five male children with insulin-dependent diabetes mellitus (IDDM) and five healthy male children. Each subject performed both types of exercise. In the healthy children, growth hormone levels rose, and glucose, glucagon, cortisol, and ketone bodies remained unchanged during both continuous and intermittent exercise. In the IDDM subjects, the mean reductions in glucose concentration were 99 mg% and 84 mg% during continuous and intermittent exercise, respectively. The levels of cortisol and ketone bodies in the IDDM subjects were significantly (P less than 0.05) elevated above the values obtained in the healthy subjects irrespective of the type of exercise. The mean concentrations of growth hormone, glucagon, cortisol, and ketone bodies were not significantly different between continuous and intermittent exercise. The study concludes that in healthy children the hormonal and metabolic responses to a continuous and an intermittent exercise protocol are similar. In the IDDM subjects, however, both forms of physical activity are associated with a decline in plasma glucose and no significant differences in hormonal and metabolic responses.


Dendrimers in layer-by-layer assemblies: synthesis and applications.
We review the synthesis of dendrimer-containing layer-by-layer (LbL) assemblies and their applications, including biosensing, controlled drug release, and bio-imaging. Dendrimers can be built into LbL films and microcapsules by alternating deposition of dendrimers and counter polymers on the surface of flat substrates and colloidal microparticles through electrostatic bonding, hydrogen bonding, covalent bonding, and biological affinity. Dendrimer-containing LbL assemblies have been used to construct biosensors, in which electron transfer mediators and metal nanoparticles are often coupled with dendrimers. Enzymes have been successfully immobilized on the surface of electrochemical and optical transducers by forming enzyme/dendrimer LbL multilayers. In this way, high-performance enzyme sensors are fabricated. In addition, dendrimer LbL films and microcapsules are useful for constructing drug delivery systems because dendrimers bind drugs to form inclusion complexes or the dendrimer surface is covalently modified with drugs. Magnetic resonance imaging of cancer cells by iron oxide nanoparticles coated with dendrimer LbL film is also discussed.


c-Jun, a crucial molecule in metastasis of breast cancer and potential target for biotherapy.
Human breast cancer cell line SKBR3 expresses high level of the ErbB2 molecule, which has been associated with poor prognosis of breast cancer. Elevated ErbB2 functions as a transactivating co-receptor and promotes the formation of ErbB2 containing heterodimers, which are more mitogenic and transforming, and have a higher ligand affinity and signaling potency by virtue of the potent latent kinase activity of ErbB2. Interestingly, SKBR3 cells are non-tumorigenic in nude mice. By ectopic overexpression of c-Jun in SKBR3 cells, involvement of c-Jun in invasiveness and metastasis in vivo was investigated in this study. The critical role of c-Jun in the tumorigenesis and metastasis is demonstrated in nude mice.


Long-term supplementation with young coconut juice does not prevent bone loss but rather alleviates body weight gain in ovariectomized rats.
Young coconut (Cocos nucifera Linn.) juice (YCJ) has traditionally been consumed to alleviate symptoms associated with the menopause. Recently, the authors demonstrated that short-term (6-week) YCJ supplementation to ovariectomized rats resulted in increased bone mass and bone formation parameter, suggesting that YCJ consumption has a positive effect on bone metabolism and may represent an intervention to help slow the bone loss during menopause transition. The present study sought to determine how long-term (12-week) YCJ supplementation affects bone metabolism in ovariectomized rats, to investigate whether such supplementation may be helpful to in osteoporosis treatment. Ten-week-old female Wistar rats were subjected to either a sham operation (Sham) or bilateral ovariectomy (Ovx). The Ovx+YCJ group received 5X-concentrated YCJ at a dose of 15 ml/kg/day for 12 weeks. Rats in the Ovx group had significantly lower femur bone mineral density than those in the Sham group. YCJ supplementation did not significantly affect this difference. However, YCJ prevented the increase in bone area of the mid third of the femur, a site high in cortical bone, and body weight gain observed following Ovx. Our findings indicate that long-term YCJ intake does not alter bone loss, but rather alleviates body weight gain following menopause.


Adverse parenting is associated with blunted salivary cortisol awakening response and altered expression of glucocorticoid receptor β and β2-adrenergic receptor mRNAs in leukocytes in Japanese medical students.
Adverse parenting is associated with an increased risk for the development of mood and behavioral disorders. In this study, we assessed the perceived parental bonding of 232 medical students using the parental bonding instrument (PBI) and extracted 22 students who reported their parents' rearing attitudes as affectionless control (LOW; low care, high overprotection). Using the 28-item general health questionnaire, the Zung self-rating depression scale (Zung-SDS), the hospital anxiety and depression scale (HADS), and the Spielberger state-trait-anxiety-inventory (STAI), physical and mental state of the LOW students were compared with those of 30 students who reported their parental bonding as optimal (OPT; high care and low overprotection). These questionnaire measurements demonstrated significantly higher anxiety and depressive mood in the LOW students versus the OPT students. Compared with the OPT students, the LOW students also exhibited a significantly reduced salivary cortisol awakening response (CAR) without changes across the rest of the diurnal salivary cortisol profile. Among glucocorticoid-related genes examined (GR, ADRB2, IκBα, IL10, IL1R2, IL1RN, MR, MC2R, TGFB1, TGFB2 and FASLG), real-time reverse transcription-PCR showed that the LOW students significantly increased expression of a dominant negative glucocorticoid receptor β (GRβ) mRNA and decreased β2-adrenergic receptor (ADRB2) mRNA levels in circulating leukocytes. These results suggest that negative perception of parents' child-rearing attitudes may be associated with anxiety and depressive mood and altered glucocorticoid signaling even in healthy young adults.


Assessment of Efficacy and Toxicity of Cyclophosphamide Chemotherapy in Canines with Malignant Mammary Tumor: A Retrospective Study.
Surgical excision with chemotherapy is a commonly used treatment modality to treat canine mammary tumor (CMT), but it is unclear whether different treatment modalities may have similar efficacies and toxicities. The objective of this clinical study was to evaluate the efficacy and toxicity of cyclophosphamide chemotherapy along with surgical excision of malignant mammary tumor in canines by clinical, haemato-biochemical, radiographical, and histopathological evaluation before and after treatment. Eighteen dogs with malignant mammary tumor, reported to Teaching Veterinary Hospital, were divided into two groups consisting of nine dogs in each group. Group I (n = 9) dogs were treated with surgical excision of malignant mammary tumor alone, and group II (n = 9) was treated with surgical excision of mammary tumor with cyclophosphamide chemotherapy at 50-100 mg/m2 intravenously in weekly doses by three consecutive weeks. In group II, 7 dogs (78%) showed complete regression of tumor after the third dose of cyclophosphamide and showed increase in the quality and survival life and remaining two dogs showed recurrence of tumor after one year. Some dogs showed common adverse reactions such as lethargy, moderate alopecia, vomiting, anorexia, anemia, and haematuria after the third dose of chemotherapy. To conclude, surgical excision combined with cyclophosphamide chemotherapy is an effective protocol for management of malignant mammary tumor in canines with minimal toxicity and it could be possible to increase the quality and survival life of patients.


[Antenatal palliative plan following diagnosis of fetal lethal condition: Rennes Teaching Hospital experience].
To study the pregnancies followed at Rennes University Hospital from 2006 to 2012, after prenatal diagnosis of lethal fetal condition and prenatal project of palliative care at birth consisting of comfort care emphasizing parent-child encounters and bonding. Retrospective study of 20 pregnancies with diagnosis of lethal fetal condition where parents accepted antenatally the proposal or sought for palliative care at birth. Diagnosis was made at a median age of 20 weeks gestation (12-33). Birth occurred at 37.4 WG, 6 caesarean sections were performed for maternal conditions. Six cases of hypoplastic left heart syndrome (HLHS) share common characteristics: good Apgar score, prolonged survival (26hours to 159days) transfer to neonatology ward (6) or later at home (4). In four multiple pregnancies, the choice of SP mainly contributed to protect healthy twins during pregnancy. In birth room, there was no need for invasive procedure or drugs. Death: one occurred during labor, 8 in birth room before H2, others in neonatal ward before d4 (excluding HLHS). These data will enable better antenatal preparation of both teams and parents. Lifetime, however short, allowed parents to meet with their child alive this permitting collection of memory traces and bonding.


In situ amplification of intracellular microRNA with MNAzyme nanodevices for multiplexed imaging, logic operation, and controlled drug release.
MicroRNAs (miRNAs), as key regulators in gene expression networks, have participated in many biological processes, including cancer initiation, progression, and metastasis, indicative of potential diagnostic biomarkers and therapeutic targets. To tackle the low abundance of miRNAs in a single cell, we have developed programmable nanodevices with MNAzymes to realize stringent recognition and in situ amplification of intracellular miRNAs for multiplexed detection and controlled drug release. As a proof of concept, miR-21 and miR-145, respectively up- and down-expressed in most tumor tissues, were selected as endogenous cancer indicators and therapy triggers to test the efficacy of the photothermal nanodevices. The sequence programmability and specificity of MNAzyme motifs enabled the fluorescent turn-on probes not only to sensitively profile the distributions of miR-21/miR-145 in cell lysates of HeLa, HL-60, and NIH 3T3 (9632/0, 14147/0, 2047/421 copies per cell, respectively) but also to visualize trace amounts of miRNAs in a single cell, allowing logic operation for graded cancer risk assessment and dynamic monitoring of therapy response by confocal microscopy and flow cytometry. Furthermore, through general molecular design, the MNAzyme motifs could serve as three-dimensional gatekeepers to lock the doxorubicin inside the nanocarriers. The drug nanocarriers were exclusively internalized into the target tumor cells via aptamer-guided recognition and reopened by the endogenous miRNAs, where the drug release rates could be spatial-temporally controlled by the modulation of miRNA expression. Integrated with miRNA profiling techniques, the designed nanodevices can provide general strategy for disease diagnosis, prognosis, and combination treatment with chemotherapy and gene therapy.