Insights on the Impact of Arbuscular Mycorrhizal Symbiosis on Eucalyptus grandis Tolerance to Drought Stress.
Drought stress has a negative impact on plant growth and production. Arbuscular mycorrhizal (AM) fungi, which establish symbioses with most terrestrial vascular plant species, play important roles in improving host plant mineral nutrient acquisition and resistance to drought. However, the physiological and molecular regulation mechanisms occurring in mycorrhizal Eucalyptus grandis coping with drought stress remain unclear. Here, we studied the physiological changes and mitogen-activated protein kinase (MAPK) cascade gene expression profiles of E. grandis associated with AM fungi under drought stress. The results showed that colonization by AM fungi significantly enhanced plant growth, with higher plant biomass, shoot height, root length, and relative water content (RWC) under drought conditions. Mycorrhizal plants had lower levels of accumulation of proline, malondialdehyde (MDA), H2O2, and O2·- than seedlings not colonized with AM fungi. In addition, mycorrhizal E. grandis also had higher peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) activities under drought conditions, improving the antioxidant system response. Eighteen MAPK cascade genes were isolated from E. grandis, and the expression levels of the MAPK cascade genes were positively induced by symbiosis with AM fungi, which was correlated with changes in the proline, MDA, H2O2, and O2·- contents and POD, SOD, and CAT activities. In summary, our results showed that AM symbiosis enhances E. grandis drought tolerance by regulating plant antioxidation abilities and MAPK cascade gene expression. IMPORTANCE Arbuscular mycorrhizal (AM) fungi play an important role in improving plant growth and development under drought stress. The MAPK cascade may regulate many physiological and biochemical processes in plants in response to drought stress. Previous studies have shown that there is a complex regulatory network between the plant MAPK cascade and drought stress. However, the relationship between the E. grandis MAPK cascade and AM symbiosis in coping with drought remains to be investigated. Our results suggest that AM fungi could improve plant drought tolerance mainly by improving the antioxidant ability to protect plants from reactive oxygen species (ROS) and alleviate oxidative stress damage. The expression of the MAPK cascade genes was induced in mycorrhizal E. grandis seedlings under drought stress. This study revealed that MAPK cascade regulation is of special significance for improving the drought tolerance of E. grandis. This study provides a reference for improving mycorrhizal seedling cultivation under stress.


3D Construction of Shape-Controllable Tissues through Self-Bonding of Multicellular Microcapsules.
Designed microtissues that replicate highly ordered three-dimensional (3D) multicellular in vivo structures have shown huge potential in biomedical research and drug discovery. Through microencapsulation and microfluidic techniques, cell-laden microcapsules have been widely used as pathological or pharmacological models. However, most conventional microtissue construction strategies can only engineer simply predefined microcapsules with monotonous biological components in two dimensions. Here, we propose a flexible 3D microtissue construction method through self-bonding of real-time shape-programmable microcapsules. The microcapsules are prepared by photo-induced electrodeposition of cell-laden alginate hydrogel and flexibly tailored into tissue-specific shapes, sizes, and arbitrary biocomponents. With the local fluidics-guided assembly, the microcapsules are spatially organized into 3D perfectly aligned microtissues. To mimic in vivo intercellular connection, the aligned microcapsules are precoated with fibroblasts to self-bond the adjacent layers into a robust assemblage through fibroblast-extracellular matrix interactions, which highly reproduces the tissue morphogenesis in natural organisms. As a typical complex tissue model, the 3D hepatic lobule was engineered utilizing HepG2 cells seeded into microcapsules with a fibroblast coating, and its biofunction including albumin and urea secretion was improved by nearly two-fold compared with cells seeded without a fibroblast coating. We anticipate that our method will be capable of regenerating more complex multicellular constructs with unprecedented possibilities for future tissue engineering applications.


[Quantitative Analysis of Immuno-fluorescence of Nuclear Factor-κB Activation].
Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/ p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.


Monitoring inflammation and airway remodeling by fluorescence molecular tomography in a chronic asthma model.
Asthma is a multifactorial disease for which a variety of mouse models have been developed. A major drawback of these models is represented by the transient nature of the airway pathology peaking 24-72 h after challenge and resolving in 1-2 weeks. We characterized the temporal evolution of pulmonary inflammation and tissue remodeling in a recently described mouse model of chronic asthma (8 week treatment with 3 allergens: Dust mite, Ragweed, and Aspergillus; DRA). We studied the DRA model taking advantage of fluorescence molecular tomography (FMT) imaging using near-infrared probes to non-invasively evaluate lung inflammation and airway remodeling. At 4, 6, 8 or 11 weeks, cathepsin- and metalloproteinase-dependent fluorescence was evaluated in vivo. A subgroup of animals, after 4 weeks of DRA, was treated with Budesonide (100 µg/kg intranasally) daily for 4 weeks. Cathepsin-dependent fluorescence in DRA-sensitized mice resulted significantly increased at 6 and 8 weeks, and was markedly inhibited by budesonide. This fluorescent signal well correlated with ex vivo analysis such as bronchoalveolar lavage eosinophils and pulmonary inflammatory cell infiltration. Metalloproteinase-dependent fluorescence was significantly increased at 8 and 11 weeks, nicely correlated with collagen deposition, as evaluated histologically by Masson's Trichrome staining, and airway epithelium hypertrophy, and was only partly inhibited by budesonide. FMT proved suitable for longitudinal studies to evaluate asthma progression, showing that cathepsin activity could be used to monitor inflammatory cell infiltration while metalloproteinase activity parallels airway remodeling, allowing the determination of steroid treatment efficacy in a chronic asthma model in mice.


Distinguishing L from M photopigment coding sequences by hybridization to novel locked nucleic acid (LNA) oligonucleotide probes.
Many attempts have been made over the years to distinguish human and primate L (long-wavelength sensitive) from M (middle-wavelength sensitive) cone photoreceptors using either immunohistochemistry or in situ hybridization. These attempts have been unsuccessful due to the very high degree of identity between the sequences of the L and M proteins and encoding mRNAs. The recent development of chemically modified oligonucleotide probes, referred to as locked nucleic acid (LNA) probes, has shown that they hybridize with much greater affinity and specificity to the target nucleic acid. This has greatly increased the potential for differentiating L from M cones by in situ hybridization. We have designed LNA oligonucleotide probes that are complementary to either the L or M coding sequences located in exon 5 of the Macaca nemestrina L and M pigment genes. We have shown that the LNA-M and LNA-L probes hybridize specifically to their respective target nucleic acid sequences in vitro. This result strongly suggests that these probes would be instrumental in rapidly distinguishing L from M cone in the entire retina, and in defining the cone mosaic during development and in adults.


[Evaluation of a direct serologic card agglutination test for the diagnosis of camel trypanosomiasis caused by Trypanosoma evansi].
The results of a novel direct serological card agglutination test for the diagnosis of camel trypanosomosis due to Trypanosoma evansi (CATT/T. evansi) were compared with those obtained by direct detection of parasites in a study using 1,093 sera from camels raised in northern Mali. A good correlation was revealed between the percentage of positive results obtained by CATT and the presence of trypanosomes (89%), as well as a good coincidence between the percentage of positive results obtained by CATT and low haematocrit values (packed cell volume). CATT revealed a global serological prevalence of 30.6%, whereas trypanosomes were found in only 5.85% of the corresponding animals. CATT/T. evansi is a quick and easy-to-read test, which merits further evaluation in camel-rearing countries.


Identifying Needs in Young Adults With Inflammatory Bowel Disease: A Qualitative Study.
Successful transitional care for young adults entails that healthcare teams recognize and understand the attitudes, perspectives, and developmental maturity of young adults. The aim of this study was to identify the needs of young adults with inflammatory bowel disease. Young adults 18-30 years of age were recruited from the McMaster University Medical Centre adult inflammatory bowel disease (IBD) clinic between July 2012 and May 2013. Semistructured interviews were audio taped, transcribed verbatim, and coded using a constant comparative method. QSR NVivo10 software was used to manage the data. Twenty-one young adults, including 15 subjects diagnosed as adolescents (younger than 18 years) and 6 subjects diagnosed as young adults, were interviewed. Four broad categories of needs were identified: psychosocial, informational, self-advocacy, and daily living needs. The most commonly reported needs were psychosocial and the least common were daily living needs. Results from this study may be used to inform clinical practitioners of potential needs that may be important to the overall quality of patient health during young adulthood. In addition, these findings may be used to evaluate existing transition and self-management tools to measure success of transition interventions more effectively.


Embryonic Coat of the Grass Shrimp Palaemonetes pugio.
The embryo of the grass shrimp, Palaemonetes pugio, is surrounded during development by a protective extracellular coat designated as the embryonic coat (EC). At hatching, this EC is composed of four embryonic envelopes (EE), each of which is composed of multiple layers. The outermost layer of the EC, the outer investment coat (OIC), is derived primarily, if not completely, from pleopods of the female. The first envelope (EE1) forms as a bilayered envelope, EE1a and EE1b, immediately after oviposition. The OIC becomes closely associated with EE1 and remains in close contact with EE1 until hatching occurs. An additional layer, EE1c, is added to the inner side of EE1 between 3 and 5 d after oviposition. Three more embryonic envelopes, EE2, EE3, and EE4, are formed between the embryo and EE1 by 7 d after oviposition. Formation of embryonic envelopes continues until 10 d after oviposition; by this time each envelope is morphologically distinct in composition, with "outer" and "inner" sides clearly identifiable. All but the innermost embryonic envelope (EE4) are shed by the embryo about 6 h before hatching. Permeability of the EC during the 12-d incubation period is found to decrease between 0 and 5 d after oviposition, and then increase until hatching. Fluorescently labeled lectins react positively with the OIC, indicating the presence of glucose and N-acetylglucosamine residues. Thus, the palaemonid EC is a dynamic structure throughout embryonic development.


Impact of a preventive intervention for perinatal depression on mood regulation, social support, and coping.
Perinatal depression prevention trials have rarely examined proximal outcomes that may be relevant for understanding long-term risk for depression. The Mothers and Babies (MB) Course is a cognitive-behavioral depression prevention intervention, which has been shown to prevent depressive symptoms among at-risk perinatal women of color. This study examined intervention impact on three proximal outcomes that are theoretically linked with the intervention's model of change and have been empirically linked with risk for depression: mood regulation expectancies, perceived social support, and coping. The study used data from a randomized intervention trial of the MB Course with 78 low-income, predominantly African-American perinatal women enrolled at one of four home visitation programs in Baltimore City. Mood regulation expectancies, perceived social support, and coping were assessed with self-report instruments at baseline, post-intervention, and 3- and 6-month follow-ups. The intervention group experienced 16 % greater growth in mood regulation from baseline to 6-month follow-up compared to the usual care group, suggesting a prevention effect. The pattern of findings was similar, although not statistically significant, for social support. Contrary to prediction, the control group experienced less growth in avoidant coping than the intervention group. Findings indicate the MB Course enhances mood regulation, which may facilitate prevention of depression over time. Assessment of intervention effects on proximal outcomes is beneficial for understanding how interventions may enhance protective factors relevant to successful long-term outcomes.


T7 RNA polymerase translocation is facilitated by a helix opening on the fingers domain that may also prevent backtracking.
Here, we studied the complete process of a viral T7 RNA polymerase (RNAP) translocation on DNA during transcription elongation by implementing extensive all-atom molecular dynamics (MD) simulations to construct a Markov state model (MSM). Our studies show that translocation proceeds in a Brownian motion, and the RNAP thermally transits among multiple metastable states. We observed non-synchronized backbone movements of the nucleic acid (NA) chains with the RNA translocation accomplished first, while the template DNA lagged. Notably, both the O-helix and Y-helix on the fingers domain play key roles in facilitating NA translocation through the helix opening. The helix opening allows a key residue Tyr639 to become inserted into the active site, which pushes the RNA-DNA hybrid forward. Another key residue, Phe644, coordinates the downstream template DNA motions by stacking and un-stacking with a transition nucleotide (TN) and its adjacent nucleotide. Moreover, the O-helix opening at pre-translocation (pre-trans) likely resists backtracking. To test this hypothesis, we computationally designed mutants of T7 RNAP by replacing the amino acids on the O-helix with counterpart residues from a mitochondrial RNAP that is capable of backtracking. The current experimental results support the hypothesis.


Meta-analysis of gemcitabine in brief versus prolonged low-dose infusion for advanced non-small cell lung cancer.
To evaluate the efficacy and safety of gemcitabine (GEM) at 30 min standard-dose infusion (30 min-SDI) compared with prolonged low-dose infusion (P-LDI) in patients with advanced non-small-cell lung cancer (NSCLC). Electronic databases including Pubmed, EMbase, Cochrane Library, CNKI, CBM, and VIP were searched using keywords "GEM", "P-LDI", and "NSCLC". Review Manager 5.3 was used to perform the meta-analysis. Primary endpoints were overall response rate (ORR) and 1-year survival rate (1-year SR). Secondary endpoints were grade 3/4 hematotoxicity and nausea/vomiting. In association. GRADE quality of evidence system was used to assess the results of meta-analysis. Six randomized controlled trials (RCTs) with a total of 637 patients were included and no statistical heterogeneity was found among the studies. The results showed that P-LDI was superior in ORR (RD = 0.09, 95% CI: 0.02 to 0.16, P = 0.02), but had a similar 1-year SR (RD = 0.05, 95% CI: -0.02 to 0.12, P = 0.18) as compared with 30 min-SDI. For grade 3/4 adverse events, there was no significant difference in anemia (RD = 0.02, 95% CI: -0.01 to 0.04, P = 0.27) and nausea/vomiting (RD = 0.01, 95% CI: -0.04 to 0.06, P = 0.64) between the two treatments. However, patients with P-LDI experienced less leukopenia (RD = -0.08, 95% CI: -0.15 to -0.01, P = 0.03) and thrombocytopenia ((RD = -0.05, 95% CI: -0.09 to -0.01, P = 0.006). The GRADE profile showed that the included RCTs had low quality of evidences. P-LDI was superior in terms of ORR, experienced less grade 3/4 thrombocytopenia and leukopenia compared with 30 min-SDI, and could be a viable treatment option for advanced NSCLC. However, the results need to be further verified by high quality trials and large samples owing to the low quality of evidences.


The pathogenesis of pulmonary edema in acute pancreatitis.
Acute pulmonary edema appeared 3 or more days after the onset of acute pancreatitis in 7 patients, an approximate incidence of 8%. The severity of pancreatitis in these patients was characterized by massive requirements for intravenous colloid and by marked hypocalcemia. In addition, at least 5 of the 7 patients had very high serum levels of triglycerides at the time of hospital admission. Hemodynamic studies during pulmonary edema showed normal central venous pressure, pulmonary artery pressure, pulmonary capillary wedge pressure, and pulmonary vascular resistance. Cardiac index was appropriately elevated. Respiratory treatment, consisting of endotracheal intubation and controlled ventilation with PEEP, was successful in allowing reversal of the pulmonary injury and recovery of respiratory function within 1-2 weeks in all cases. Two patients died later from pancreatic abscesses. The findings indicate that a distinct form of pulmonary injury may occur in acute pancreatitis, characterized by loss of integrity of the alveolar-capilllary membrane, leading to pulmonary edema. The mechanism of injury is not known but may be caused by circulating free fatty acids, phospholipase A, or vasoactive substances. The pulmonary membrane lesion appears to heal during the period of intensive respiratory support.


Conformational substates modulate hydride transfer in dihydrofolate reductase.
In earlier studies of the hydride-transfer reaction catalyzed by dihydrofolate reductase (DHFR) we identified features of the protein correlated with variations in the reaction barrier. We extend the scope of those studies by carrying out potential of mean force (PMF) simulations to determine the hydride-transfer barrier in the wild-type protein as well as the G121V and G121S mutants. While our prior studies focused on the reactant state, our current work addresses the full reaction pathway and directly probes the reactive event. The free energy barriers and structural ensembles resulting from these PMF calculations exhibit the same trends reported in our previous work. Fluctuations present in these simulations also exhibit trends associated with differences in the hydride-transfer barrier height. Moreover, vibrational modes anticipated to promote hydride transfer exhibit larger amplitudes in simulations that generate lowered barriers. The results of our study indicate that discrete basins (substates) on a potential energy landscape of the enzyme give rise to distinct hydride-transfer barriers. We suggest that the long-range effects of mutations at position 121 within DHFR are mediated by differentially preorganized protein environments in the context of distinct substate distributions, with concomitant changes to the dynamic properties of the enzyme.


Multiple histone deacetylases and the CREB-binding protein regulate pre-mRNA 3'-end processing.
Trichostatin A (TSA), a specific inhibitor of histone deacetylases (HDACs), induces acetylation of various non-histone proteins such as p53 and alpha-tubulin. We purified several acetylated proteins by the affinity to an anti-acetylated lysine (AcLys) antibody from cells treated with TSA and identified them by mass spectrometry. Here we report on acetylation of CFIm25, a component of mammalian cleavage factor Im (CF Im), and poly(A) polymerase (PAP), a polyadenylating enzyme for the pre-mRNA 3'-end. The residues acetylated in these proteins were mapped onto the regions required for interaction with each other. Whereas CBP acetylated these proteins, HDAC1, HDAC3, HDAC10, SIRT1, and SIRT2 were involved in in vivo deacetylation. Acetylation of the CFIm25 occurred depending on the cleavage factor complex formation. Importantly, the interaction between PAP and CF Im complex was decreased by acetylation. We also demonstrated that acetylation of PAP inhibited the nuclear localization of PAP by inhibiting the binding to the importin alpha/beta complex. These results suggest that CBP and HDACs regulate the 3'-end processing machinery and modulate the localization of PAP through the acetylation and deacetylation cycle.


[Effect of urotensin II on iNOS expression in human umbilical vein endothelial cells].
To observe the direct effect of urotensin II (U II) on the release of nitric oxide (NO) and expression of inducible nitric oxide synthase (iNOS) mRNA in human umbilical vein endothelial cells (HUVEC). HUVEC were cultured with different concentrations of U II (10(-9)-10(-7) mol/L) for 24 hours. Then the supernatant was collected to detect the level of NO and the activity of iNOS, the expression of iNOS mRNA of HUVEC was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). In comparison with controls, the level of NO, the activity of iNOS and the iNOS mRNA expression increased significantly (P < 0.05). U II may up-regulate the expression of iNOS mRNA and increase NO generation in HUVEC, it suggests that U II may relax blood vessel by activating iNOS/NO pathway.


Time-Resolved Small-Angle X-ray Scattering Reveals Millisecond Transitions of a DNA Origami Switch.
Self-assembled DNA structures enable creation of specific shapes at the nanometer-micrometer scale with molecular resolution. The construction of functional DNA assemblies will likely require dynamic structures that can undergo controllable conformational changes. DNA devices based on shape complementary stacking interactions have been demonstrated to undergo reversible conformational changes triggered by changes in ionic environment or temperature. An experimentally unexplored aspect is how quickly conformational transitions of large synthetic DNA origami structures can actually occur. Here, we use time-resolved small-angle X-ray scattering to monitor large-scale conformational transitions of a two-state DNA origami switch in free solution. We show that the DNA device switches from its open to its closed conformation upon addition of MgCl2 in milliseconds, which is close to the theoretical diffusive speed limit. In contrast, measurements of the dimerization of DNA origami bricks reveal much slower and concentration-dependent assembly kinetics. DNA brick dimerization occurs on a time scale of minutes to hours suggesting that the kinetics depend on local concentration and molecular alignment.


Costs for 'hospital at home' patients were 19 percent lower, with equal or better outcomes compared to similar inpatients.
Hospitals are the standard acute care venues in the United States, but hospital care is expensive and can pose health threats for older people. Albuquerque, New Mexico-based Presbyterian Healthcare Services adapted the Hospital at Home® model developed by the Johns Hopkins University Schools of Medicine and Public Health to provide acute hospital-level care within patients' homes. Patients show comparable or better clinical outcomes compared with similar inpatients, and they show higher satisfaction levels. Available to Medicare Advantage and Medicaid patients with common acute care diagnoses, this program achieved savings of 19 percent over costs for similar inpatients. These savings were predominantly derived from lower average length-of-stay and use of fewer lab and diagnostic tests compared with similar patients in hospital acute care. Hospital at Home advances the Triple Aim of clinical quality, affordability, and exceptional patient experience.


[Surveillance on hemorrhagic fever with renal syndrome in Guizhou during 1984-2000].
To explore the relations between different areas on hemorrhagic fever with renal syndrome (HFRS) in Guizhou. Various forms of infectious areas were surveyed continuously through epidemiologic surveillance system. In fixed areas under surveillance system, the mean positive rate of HFRSV among Apodemus agrarius was 3.39%, comparing with Rattus norvegicus 1.61% in Apodemus infectious areas of Zunyi county, 3.19% in Rattus norvegicus, but no HFRSV of Apodemus agrarius was identified in Rattus infectious area of Shiqian county. Both Apodemus and Rattus infectious areas were relatively stabilized. In both banks of Luowang river, Kaiyang county, which had been identified as areas of infections for Apodemus in the eastern part, Rattus infectious area in the west, slow change was noticed. In 1983 - 1984 was not found in Apodemus agrarius HFRSV, however the infectious rate of HFRSV in Apodemus agrarius was 13.85% (Ag 1/65, Ab 8/65) in the western part of the province in 1995 - 1998. Both Apodemus and Rattus infectious areas were stabilized but changed slowly. Mixed type and the result of mutual penetration were noticed.


Inhalation treatment for asthma.
Inhaled medication has revolutionised the lives of many children with asthma. Despite this we see many children for whom appropriate inhaled medication has been prescribed but whose symptoms continue to be poorly controlled. In our experience this is often due to poor technique or inappropriately prescribed devices and an inadequate understanding of when and how to use the treatment. The prescribing physician must have a clear idea of the optimal inhalation technique. We have reviewed the standard devices available and our use of them in the treatment of childhood asthma.


Giant arachnoid granulation: differential diagnosis of acute headache.
We present a case of intense, rapidly evolving headache clinically mimicking meningitis, subarachnoid haemorrhage or venous sinus thrombosis. Clinical examination, standard blood work and central nervous system studies were non-contributory and effectively ruled out these diagnoses. Cranial multidetector CT studies before and after application of intravenous contrast medium performed prior to lumbar tap disclosed a non-enhancing ovoid mass filling the superior sagittal sinus. This lesion posed a differential to venous sinus thrombosis, but ultimately fulfilled the criteria of a giant arachnoid granulation. The imaging characteristics and differential diagnosis of giant arachnoid granulations are discussed.


Reliability of measuring facial morphology with a 3-dimensional laser scanning system.
The purpose of this prospective clinical trial was to evaluate the reliability of a 3-dimensional facial scanning technique for the measurement of facial morphology. A field study was conducted in 2 comprehensive schools in the South Wales region of the United Kingdom. Forty subjects, mean age 11 years 3 months, were analyzed for soft tissue changes at baseline (T1), within 3 minutes (T2), and 3 days later (T3) by using 2 commercially available Minolta Vivid 900 (Osaka, Japan) laser-scanning devices assembled as a stereo pair. Left and right images were merged to form the whole face, and these images were superimposed to assess the errors at T1 and T2, and T1 and T3. The results showed that premerged left and right mean shell deviations were 0.38 +/- 0.14 mm for scans at T1, 0.31 +/- 0.09 mm at T2, and 0.34 +/- 0.12 mm at T3. The mean differences of the merged composite face were 0.31 +/- 0.08 mm between T1 and T2, and 0.40 +/- 0.11 mm between T1 and T3. Paired t tests showed no significant difference between these groups (P > .05). Shell deviation facial maps of the merged scans showed that 90% of the created composite facial scans were within an error of 0.85 mm. Capturing the soft tissue morphology of the face with this technique is clinically reproducible within 3 minutes and 3 days of the initial records.


Simultaneous determination of procymidone, pyridaben and beta-cypermethrin residues in tea solution by GC-ECD.
A sensitive and effective method for the simultaneous quantitative determination of procymidone, pyridaben and beta-cypermethrin residues in tea solutions was developed and validated using a gas chromatograph (GC) equipped with an electron capture detector (ECD). The analysis of the three pesticides in tea solutions involved extraction with petroleum ether-ethyl acetate (3:1, v/v), cleanup using a Florisil solid-phase extraction cartridge and subsequent determination by GC-ECD. Recovery studies were carried out at three spiked levels (0.05, 0.1 and 0.5 mg/kg). The overall average recoveries using this method in green teas and black teas at the three concentration levels ranged from 85.63 to 105.80% with relative standard deviations in the range of 1.29-4.97% (n = 5) for all analytes. The quantification limits of procymidone, pyridaben and beta-cypermethrin were 0.025, 0.038 and 0.030 mg/kg, respectively, which were lower than the maximum residue limits (MRLs) of 0.1 mg/kg procymidone, 0.05 mg/kg pyridaben and 0.5 mg/kg beta-cypermethrin in tea samples established by European Union legislations. This study provides a theoretical basis for China to draw up MRLs for procymidone, pyridaben and beta-cypermethrin in tea solutions.


Effect of moisture on disintegration kinetics during anaerobic digestion of complex organic substrates.
The role of the moisture content and particle size (PS) on the disintegration of complex organic matter during the wet anaerobic digestion (AD) process was investigated. A range of total solids (TS) from 5% to 11.3% and PS from 0.25 to 15 mm was evaluated using carrot waste as model complex organic matter. The experimental results showed that the methane production rate decreased with higher TS and PS. A modified version of the AD model no.1 for complex organic substrates was used to model the experimental data. The simulations showed a decrease of the disintegration rate constants with increasing TS and PS. The results of the biomethanation tests were used to calibrate and validate the applied model. In particular, the values of the disintegration constant for various TS and PS were determined. The simulations showed good agreement between the numerical and observed data.


Decontamination of Creutzfeldt-Jakob disease and other transmissible agents.
The bovine spongiform encephalopathy (BSE) epidemic in cows, and the recent BSE-linked human infections, present new public health problems. More rigorous measures are needed to prevent additional transmissions. Tissue from established but undiagnosed human infections can contaminate medical supplies and instruments. We tested guanidine thiocyanate (GdnSCN) solutions and found them to be highly effective in disrupting the infectious agent, even in very complex tissues such as whole brain. It may be prudent now to use this reagent routinely in surgical and other relevant settings.


Pollution induced nitrative stress and heat shock protein 70 overexpression in fish liver mitochondria.
Exposure to heavy metals and organic pollutants in natural water bodies can have detrimental effects on fish health. A combination of biochemical and energy studies were used to observe the changes in fish liver mitochondria in response to environmental pollutant induced nitrative stress in natural field conditions. The fish samples Mugil cephalus were collected from polluted (Ennore) and unpolluted (Kovalam) estuaries for a period of two years. The results revealed elevated nitrite (NO2-) and nitrate (NO3-) levels, increased nitric oxide (NO) synthesis and 3-nitrotyrosine expression, decreased respiratory chain enzyme activities and ATP/ADP ratio, reduced mitochondrial superoxide dismutase (MnSOD), glutathione peroxidase (Gpx) levels, diminished thiol status that leads to alterations in the mitochondrial function and elevated mitochondrial heat shock protein 70 (mtHSP70) expression (30%) to a significant extent in fish from the polluted estuary than in the unpolluted estuary. The overexpression of HSP70 under stress may aid mitochondrial survival by protecting against nitrative stress induced damage. The results also reveal the percentage increase in fish liver mitochondrial HSP70 in response to cumulative effect of environmental pollutants.


Nonspecific cleavage of phi X174 RFI deoxyribonucleic acid by bleomycin.
The covalently closed circular duplex deoxyribonucleic acid (DNA) of phi X174 underwent progressive conversion to nicked and linear DNA with increasing bleomycin/phi X174 RFI DNA molecule ratios. The formation of linear DNA (a double-strand break) occurred under limited reaction conditions as low as an average of 0.2 single-strand break/phi X174 RFI DNA molecule. As bleomycin-produced linear DNA was further fragmented by bleomycin, a broad distribution of DNA fragments without notable concentrations of unique size was formed. Restriction enzymes PstI and SstII did not generate discrete fragments from bleomycin-produced full-length linear phi X174 DNA, nor did bleomycin cleavage generate discrete fragments from HpaII or PstI digests of phi X174 RFI. These findings suggest that bleomycin does not act at a few specific sites on phi X174 RFI DNA. The single-strand nick appeared to be the preferred site for bleomycin action for a second cleavage in a phi X174 molecule.


The Holy Grail of Polymer Therapeutics for Cancer Therapy: An Overview on the Pharmacokinetics and Bio Distribution.
In recent years, multifaceted clinical benefits of polymeric therapeutics have been reported. Over the past decades, cancer has been one of the leading causes of mortality in the world. Many clinically approved chemotherapeutics encounter potential challenges against deadly cancer. Moreover, safety and efficacy of anticancer agents have been limited by undesirable pharmacokinetics and biodistribution. To address these limitations, various polymer drug conjugates are being studied and developed to improve the antitumor efficacy. Among other therapeutics, polymer therapeutics are well established platforms that circumvent anticancer therapeutics from enzymatic metabolism via direct conjugation to therapeutic molecules. Interestingly, polymer therapeutics meets an unmet need of small molecules. Further clinical study showed that polymer-drug conjugation can achieve desired pharmacokinetics and biodistribution properties of several anticancer drugs. The present retrospective review mainly enlightens the most recent preclinical and clinical studies include safety, stability, pharmacokinetic behavior and distribution of polymer therapeutics.


Augmenting effect of serotonin on the voltage-dependent Ca2+ current and subsequently activated K+ current in Aplysia neurons.
Receptor-induced activation of protein kinase C (PKC) plays an important role in modulation of various types of ionic channels in neurons. For example, PKC causes facilitation or long-lasting activation of certain ionic channels involved in spike firing after the receptor stimulation. We investigated the effect of serotonin (5-HT) on the voltage-dependent Ca(2+) channels in RB and RC neurons of Aplysia ganglia under voltage clamp. An outward current response was induced by voltage change of the cell membrane from -60 mV to +10 mV. Application of 5-HT significantly augmented the outward current response to the voltage change. Both the outward current and the augmenting effect of 5-HT markedly decreased when examined in either Ca(2+)-free, 10 mM tetraethylammonium, or 0.3 mM Cd(2+)-solution, indicating the current to be Ca(2+)-activated K(+) current produced by Ca(2+) entry. Intracellular application of either guanosine 5'-O-(2-thiodiphosphate) or cholera toxin (CTX), reagents for G-proteins, irreversibly blocked the augmenting effect of 5-HT. Application of phorbol dibutylate (PDBu), an activator of PKC, augmented the outward current and the effect of 5-HT was occluded after PDBu application. Staurosporine, a specific inhibitor of PKC, markedly suppressed the augmenting effects of both 5-HT and PDBu on the outward current. However, either 5-HT or PDBu did not augment the Ca(2+)-activated K(+) current induced by intracellular injection of Ca(2+) but rather depressed it. These results suggest that stimulation of 5-HT receptor may activate a novel type of CTX-sensitive G-protein and subsequent PKC, and that phosphorylation of voltage-dependent Ca(2+) channels may result in the increase in Ca(2+) entry and subsequent Ca(2+)-activated K(+) current. The mechanism may contribute to retain the long-lasting activation without broadening of the spike width during the excitatory response to 5-HT in these neurons.


Effect of catgut embedding at acupoints versus non-acupoints in abdominal obesity: a randomized clinical trial.
To explore the difference of catgut embedding effect between acupoints and non-acupoints in patients with abdominal obesity (AO). In this multicenter, double-blind, randomized controlled trial, all subjects were randomly assigned into the acupoint catgut embedding (ACE) group and control group (catgut embedding at non-acupoints). With a 12-week actual intervention period and a 4-week period of follow-up. Waist circumference (WC), body weight, body mass index (BMI), hip circumference (HC) and appetite were applied and assessed at baseline and after 6, 12 and 16 weeks. After the total intervention phase (12 weeks), the WC, body weight, BMI, HC and visual analogue scale scores of appetite, decreased significantly in the two groups as compared to the baseline (0.001). Meanwhile, after the 4-week follow-up, the indicators still decreased significantly in the ACE group (0.001). At 12 and 16 weeks, catgut embedding at acupoints showed significantly advantages to non-acupoints in WC and appetite (0.05). No serious adverse events were observed in ACE group and control group. Catgut embedding at acupoints and non-acupoints are all effective and safe for AO. ACE can effectively treat AO as expected and deliver lasting results.


Colour categories are reflected in sensory stages of colour perception when stimulus issues are resolved.
Debate exists about the time course of the effect of colour categories on visual processing. We investigated the effect of colour categories for two groups who differed in whether they categorised a blue-green boundary colour as the same- or different-category to a reliably-named blue colour and a reliably-named green colour. Colour differences were equated in just-noticeable differences to be equally discriminable. We analysed event-related potentials for these colours elicited on a passive visual oddball task and investigated the time course of categorical effects on colour processing. Support for category effects was found 100 ms after stimulus onset, and over frontal sites around 250 ms, suggesting that colour naming affects both early sensory and later stages of chromatic processing.


Molecular mechanisms of rosmarinic acid from Salvia miltiorrhiza in acute lymphoblastic leukemia cells.
Rosmarinic acid (RA), a major hydrosoluble bioactive compound found in the Chinese medicinal herb, Salvia miltiorrhiza Bunge, which has been used in traditional Chinese medicine to treat various diseases, including cancer. However, the mechanisms have not been fully elucidated. Guided by microarray hybridization and Ingenuity Pathway Analysis, we identified modes of action of rosmarinic acid (RA) isolated from S. miltiorrhiza on acute lymphoblastic leukemia cells. Microarray data were verified by independent methods: Real-time RT-PCR (mRNA expression), resazurin assay (cytotoxicity of RA towards parental CCRF-CEM, multidrug-resistant CEM/ADR5000 cells and normal lymphocytes), flow cytometry (cell cycle arrest, apoptosis, necroptosis, generation of reactive oxygen species (ROS), disruption of mitochondrial membrane potential (MMP)), single cell gel electrophoresis (DNA damage), molecular docking and gene promoter binding motif analysis (NFκB), Western blotting (nuclear NFκB translocation, PARP cleavage, caspase 3/7/9 expression), and fibronectin-based cell adhesion assay. RA dose-dependently inhibited CCRF-CEM and CEM/ADR5000 cells, but caused less cytotoxicity towards normal lymphocytes. RA simultaneously induced apoptosis and necrosis, as shown by cell morphology and annexin V-PI assay. DNA damage was dose-dependently induced without ROS generation, which subsequently led to cell cycle arrest. RA-stimulated MMP dysfunction activated PARP-cleavage and caspase-independent apoptosis. In accordance with molecular docking and gene promoter binding motif analyses, p65 translocation from the cytosol to the nucleus was blocked by RA, indicating a mechanistic role of the NFκB pathway to explain RA's action. RA affected cellular movement as evaluated by ameliorating cell adhesion to fibronectin. RA induced apoptosis and necrosis in a ROS-independent DNA damage and caspase-independent manner. These results may contribute to the rationale use of S. miltiorrhiza and RA in traditional medicine of leukemia.


Hetero di- and trinuclear Cu-Gd complexes with trifluoroacetate bridges: synthesis, structural and magnetic studies.
The synthesis, structural determinations and magnetic properties of an original dinuclear Cu-Gd and two trinuclear Cu-Ln-Cu complexes (Ln = Gd or Dy) made with a Schiff base ligand and carboxylate ligands are reported. In each complex, the copper and gadolinium or dysprosium ions are linked by a double phenoxo bridge and a eta1:eta1:mu trifluoroacetato bridge. The dinuclear Cu-Gd complex exhibits the usual ferromagnetic interaction while the trinuclear (Cu-Gd-Cu) complexes can not be fitted with a simple Hamiltonian that only takes into account the Cu-Gd interaction. Introduction into the Hamiltonian of a Cu-Cu interaction between the terminal copper ions gives a nice fit in the case of a ferromagnetic Cu-Cu interaction (J'(Cu-Cu) = 8.3 and 9.8 cm(-1)). For the first time, the occurrence of a ferromagnetic interaction between the copper ions of the trinuclear entities is evidenced and supported by the field dependence of the magnetization. As there is no direct proof for the existence of an interaction between two centers through a third magnetic center located in between them, we can conclude that the existence of pi-pi stacking interactions through the phenyl rings of the ligands are responsible for the presence of these interactions.


ewrapper: Operationalizing engagement strategies in mHealth.
The advancement of digital technologies particularly in the domain of mobile health (mHealth) holds great promise in the promotion of health behavior. However, keeping users engaged remains a central challenge. This paper proposes a novel approach to address this issue by supplementing existing and future mHealth applications with an engagement wrapper - a collection of engagement strategies integrated into a single, coherent model. The engagement wrapper is operationalized within the format of an ambient display on the lock screen of mobile devices.


Calpain and mitochondria in ischemia/reperfusion injury.
Studies of ischemia/reperfusion (I/R) injury and preconditioning have shown that ion homeostasis, particularly calcium homeostasis, is critical to limiting tissue damage. However, the relationship between ion homeostasis and specific cell death pathways has not been investigated in the context of I/R. Previously we reported that calpain cleaved Bid in the absence of detectable caspase activation (1). In this study, we have shown that an inhibitor of the sodium/hydrogen exchanger prevented calpain activation after I/R. Calpain inhibitors prevented cleavage of Bid as well as the downstream indices of cell death, including DNA strand breaks, creatine kinase (CK) release, and infarction measured by triphenyl tetrazolium chloride (TTC) staining. In contrast, the broad spectrum caspase inhibitor IDN6734 was not protective in this model. To ascertain whether mitochondrial dysfunction downstream of these events was a required step, we utilized a peptide corresponding to residues 4-23 of Bcl-x(L) conjugated to the protein transduction domain of HIV TAT (TAT-BH4), which has been shown to protect mitochondria against Ca2+-induced deltaPsi(m) loss (2). TAT-BH4 attenuated CK release and loss of TTC staining, demonstrating the role of mitochondria and a pro-apoptotic Bcl-2 family member in the process leading to cell death. We propose the following pathway. (i) Reperfusion results in sodium influx followed by calcium accumulation. (ii) This leads to calpain activation, which in turn leads to Bid cleavage. (iii) Bid targets the mitochondria, causing dysfunction and release of pro-apoptotic factors, resulting in DNA fragmentation and death of the cell. Ischemia/reperfusion initiates a cell death pathway that is independent of caspases but requires calpain and mitochondrial dysfunction.


Combination antibiotic therapy is appropriate for nosocomial pneumonia in the intensive care unit.
Pneumonia in the intensive care unit (ICU) has been associated with highly virulent pathogens and mortality rates of 30% to 70%. Initial empirical therapy for pneumonia in the ICU remains controversial. Monotherapy with a broad-spectrum beta-lactam or fluoroquinolone may reduce toxicities associated with aminoglycosides but in nosocomial settings may fail to cover relevant pathogens and may promote antimicrobial resistance. Combining antibiotics that act by different mechanisms may achieve synergistic killing, expand the antimicrobial spectrum, and limit the emergence of antimicrobial resistance. Unfortunately, this approach may increase costs and toxicities. A plethora of studies have assessed monotherapy or combinations of antimicrobials for empirical treatment of fever in immunocompromised hosts, but these studies cannot be extrapolated to nosocomial pneumonia. Few studies have compared these disparate therapeutic strategies for nosocomial pneumonia. In this review, we emphasize the limitations of existing literature and present an approach to therapy of severe nosocomial pneumonia. Despite potential toxicities, aminoglycosides may have an important adjunctive role as empirical therapy of severe nosocomial pneumonias (pending clarification of the responsible organism) or when P aeruginosa, Acinetobacter spp, or beta-lactamase-producing aerobic gram-negative bacteria are suspected. The alarming increase in antimicrobial resistance noted in nosocomial settings in recent years, a trend that may be facilitated by extensive use of particular classes of antibiotics, provides a compelling argument in favor of combination therapy. Novel strategies employing beta-lactam and fluoroquinolone combinations also have promise.


Relationship Between Angiotensin-converting Enzyme Gene Polymorphism and QT Dispersion in Hemodialysis Patients.
The angiotensin-converting enzyme (ACE) gene insertion or deletion in long-term hemodialysis patients may be associated with corrected QT interval prolongation, leading to fatal arrhythmias. The ACE D allele is known to increase the risk of malignant ventricular arrhythmias and is also associated with increased QT dispersion after myocardial infarction and hypertension. This study aimed to evaluate the relationship between ACE gene polymorphism and QT dispersion in hemodialysis patients. In 70 hemodialysis patients, electrocardiography was performed and QT dispersion was calculated. Corrected QT interval was calculated using Bazett Formula. The ACE gene polymorphism was determined by polymerase chain reaction. The mean age of the patients was 60 ± 12 years. The mean QT dispersion and corrected QT dispersion were 61.71 ± 21.99 and 73.18 ± 25.51, respectively. QT dispersion inversely correlated with serum calcium and potassium levels and positively correlated with ACE gene polymorphism and residual urine. Calcium level was the predictor factor for QT dispersion. The ACE genotype correlated with QT dispersion, corrected QT dispersion, hemoglobin, and residual urine, and inversely correlated with serum potassium. Corrected QT dispersion correlated with ACE gene polymorphism and residual urine. The DD genotype of ACE had significally greater QT dispersion and corrected QT dispersion than the II and ID genotypes. Our study showed that the most important parameter affecting corrected QT dispersion was ACE gene polymorphism on the background of D allelle. Patients carrying this allelle need special attention regarding optimal suppression of renin-angiotensin-aldosteron system activity.


Metabolic Impact of Redox Cofactor Perturbations on the Formation of Aroma Compounds in Saccharomyces cerevisiae.
Redox homeostasis is a fundamental requirement for the maintenance of metabolism, energy generation, and growth in Saccharomyces cerevisiae. The redox cofactors NADH and NADPH are among the most highly connected metabolites in metabolic networks. Changes in their concentrations may induce widespread changes in metabolism. Redox imbalances were achieved with a dedicated biological tool overexpressing native NADH-dependent or engineered NADPH-dependent 2,3-butanediol dehydrogenase, in the presence of acetoin. We report that targeted perturbation of the balance of cofactors (NAD(+)/NADH or, to a lesser extent, NADP(+)/NADPH) significantly affected the production of volatile compounds. In most cases, variations in the redox state of yeasts modified the formation of all compounds from the same biochemical pathway (isobutanol, isoamyl alcohol, and their derivatives) or chemical class (ethyl esters), irrespective of the cofactors. These coordinated responses were found to be closely linked to the impact of redox status on the availability of intermediates of central carbon metabolism. This was the case for α-keto acids and acetyl coenzyme A (acetyl-CoA), which are precursors for the synthesis of many volatile compounds. We also demonstrated that changes in the availability of NADH selectively affected the synthesis of some volatile molecules (e.g., methionol, phenylethanol, and propanoic acid), reflecting the specific cofactor requirements of the dehydrogenases involved in their formation. Our findings indicate that both the availability of precursors from central carbon metabolism and the accessibility of reduced cofactors contribute to cell redox status modulation of volatile compound formation.


[Initial experience of catheter ablation of ventricular tachycardia originate from endocardium via direct ventricle puncture access in patients underwent mechanical valve implantation].
Objective: To evaluate the results of catheter ablation of ventricular tachycardia (VT) via direct ventricle puncture access in patients without traditional approach. Methods: Two idiopathic left fasicular VT patients with mechanical aortic and mitrial valve repalcement and 1 patient with right ventricular originated VT post mechanical tricuspid valve repalcement from March 2010 to July 2012 in Fuwai hospital were enrolled in this study. For left fasicular VT patients, catheter ablation was performed using transapical left ventricular access via minithoracotomy. For the patient with right ventricular originated VT, catheter ablation was performed via percutaneous right ventricle puncture at xiphoid. Abaltion was guided under EnSite NavX mapping system. The feasibility of VT ablation via direct ventricle puncture access and long-term VT recurrence were investigated. Results: Catheter ablation was successful in all patients, and all clinical VTs were eliminated. The procedure time was 53, 62 and 74 minutes respectively with radiation time 11, 16 and 20 minutes. The ablation time was 130, 170 and 240 seconds individually. No procedure related complication occurred. After a follow-up time of 76, 55 and 82 months respectively, no VT recurrence was found in patients with left fasicular VT. New-onset VT with different morphology with previous VT was recorded in the patient with right ventricular originated VT, subcutaneous implantable defibrillator was implanted finally in this patient. Conclusions: For patients with endocardial origined ventricular arrhythmias which could not be ablated via traditional approaches, direct ventricle puncture access with hybrid techniques provides a new approach foreliminating VTs in these patients.


Predictors of orthorexic behaviours in patients with eating disorders: a preliminary study.
The construct of orthorexia in eating disorders (EDs) has received very little attention despite clinical observations of a possible overlap between the two. The aim of this study was: 1) to assess orthorexic behaviours, eating disorder pathology and attitudinal body image in ED patients; 2) to identify possible predictors of orthorexia nervosa among ED patients. Fifty-two women diagnosed with EDs were recruited. Patients' assessment included the following: the ORTO-15 test (Polish version) for orthorexic behaviours; the Eating Attitude Test-26 (EAT-26) to identify ED symptoms; the Multidimensional Body-Self Relations Questionnaire (Polish version) to assess body image. A latent class analysis was performed and differences between identified classes were assessed. The main differences concerned weight, ED pathology and orthorexic behaviours within the same group of ED patients. In order to examine predictors of orthorexia nervosa, we investigated a structural equation model, which excellently fitted to the data (χ(2)(17) = 23.05; p = .148; CFI = .962; RMSEA = .08; p = .25; SRMR = .05). In ED patients, orthorexic behaviour was negatively predicted by eating pathology, weight concern, health orientation and appearance orientation. The assessment of the orthorexia construct in EDs may add to the paucity of studies about this issue and may help to clarify the relationship between the two. Differences and similarities seem to exist between these disorders, and may benefit from specific treatment approaches. Moreover, these preliminary findings open tracks for future research in the field of the psychology of eating.


Charge and energy dynamics in photo-excited poly(para-phenylenevinylene) systems.
We report results from simulations of charge and energy dynamics in poly(para-phenylenevinylene) (PPV) and PPV interacting with C60. The simulations were performed by solving the time-dependent Schrodinger equation and the lattice equation of motion simultaneously and nonadiabatically. The electronic system and the coupling of the electrons to the lattice were described by an extended three-dimensional version of the Su-Schrieffer-Heeger model, which also included an external electric field. Electron and lattice dynamics following electronic excitations at different energies have been simulated. The effect of additional lattice energy was also included in the simulations. Our results show that both exciton diffusion and transitions from high to lower lying excitations are stimulated by increasing the lattice energy. Also field induced charge separation occurs faster if the lattice energy is increased. This separation process is highly nonadiabatic and involves a significant rearrangement of the electron distribution. In the case of PPV coupled to C60, we observe a spontaneous charge separation. The separation time is in this case limited by the local concentration of C60 molecules close to the PPV chain.


Multiple chemical sensitivity syndrome: a clinical perspective. I. Case definition, theories of pathogenesis, and research needs.
Multiple chemical sensitivity syndrome (MCS) does not appear to fit established principles of toxicology. Yet social, political, and economic forces are demanding that MCS be defined medically, even though to date scientific studies have not identified pathogenic mechanisms for the condition or any objective diagnostic criteria. Consequently, a working definition of MCS can rely only on an individual's subjective symptoms of distress and attribution to environmental exposures rather than currently measurable objective evidence of disease. Nevertheless, patients labeled with MCS are clearly distressed and many are functionally disabled. In this review, four theories of causation are explored: (1) MCS is a purely biologic/physical or psychophysiologic reaction to low-level chemical exposures. (2) MCS symptoms may be elicited by low-level environmental chemical exposures, but the sensitivity is initiated by psychologic stress. (3) MCS is a misdiagnosis and chemical exposure is not the cause. The symptoms may be due to misdiagnosed physical or psychologic illness. (4) MCS is an illness belief system manifest by culturally shaped illness behavior. Areas for further research regarding the etiologies of MCS are suggested. Recognizing that the cause of the syndrome may be multifactorial, strategies are proposed for clinical evaluation and management in Part II of this manuscript using a biopsychosocial model of illness.


Correlation of Preoperative and Radical Prostatectomy Gleason Score: Examining the Predictors of Upgrade and Downgrade Results.
Preoperative Gleason score (GS) obtained from Trans Rectal Ultra Sonography (TRUS) is the most common grading system to evaluate the appropriate treatment for patients with clinically localized prostate cancer. But this method showed upgraded and downgraded results in comparison to Gleason score obtained from radical prostatectomy. The current study aimed to determine clinical or pathological variables to reduce the differences between biopsy and radical prostatectomy Gleason scores.Through retrospective review of 52 patients with radical prostatectomy, this study examined the correlations of preoperative Gleason score with age, prostate volume, PSA level, PSA density, digital rectal exam findings and percentage of positive core needle biopsies across two groups, including patients with preoperative GS≤6 (i.e. group one) and patients with preoperative GS≥7 (group two). The discordance between biopsy GS and radical prostatectomy GS was observed to be 52% in the current study. Among patients with preoperative GS≤6, prostate volume (P=0.026), PSA density (P=0.032) and percentage of positive core needle biopsies (P=0.042) were found to be significant predictors for upgrade. There was no significant predictor for downgrade in patients with preoperative GS≥7. Findings of this study revealed that in patients with preoperative GS≤6, smaller prostate volume, higher prostate density and higher positive results of core needle biopsies were associated with theupgrade of GS. Therefore, it should be considered when selecting treatment modalities among these patients.


Malignant proliferating trichilemmal tumor of the lower eyelid.
To report on the first case of malignant proliferating trichilemmal tumor (PTT) on the eyelid, and to describe the clinical and histopathologic findings. Interventional case report and literature review. A 42-year-old woman presented with a 2-year history of a firm, 3 x 2 cm lower eyelid mass. The mass was excised with the frozen section control. The defect in the lower eyelid was reconstructed with modified Hughes tarsoconjunctival flap and skin graft from the left postauricular area. Histopathologic examination revealed the proliferation of outer hair sheath epithelium with multiple central areas of trichilemmal keratinization. The presence of marked cellular atypia and frequent mitoses indicated the malignant transformation. Malignant PTT has not yet been reported on the eyelid. Malignant PTT should be included in the differential diagnosis for eyelid tumors.


Dietary habits and weight maintenance success in high versus low exercisers in the National Weight Control Registry.
The National Weight Control Registry (NWCR) was established to examine characteristics of successful weight loss maintainers. This study compares the diet and behavioral characteristics and weight regain trajectories of NWCR members with differing physical activity (PA) levels at baseline. Participants (n = 3591) were divided into 4 levels of self-reported PA at registry entry (< 1000, 1000 to < 2250, 2250 to < 3500, and ≥ 3500 kcals/week). We compared self-reported energy intake (EI), macronutrient composition, eating behaviors (dietary restraint, hunger, and disinhibition), weight loss maintenance strategies, and 3 year weight regain between these 4 activity groups. Those with the highest PA at registry entry had lost the most weight, and reported lower fat intake, more dietary restraint, and greater reliance on several specific dietary strategies to maintain weight loss. Those in the lowest PA category maintained weight loss despite low levels of PA and without greater reliance on dietary strategies. There were no differences in odds of weight regain at year 3 between PA groups. These findings suggest that there is not a "one size fits all strategy" for successful weight loss maintenance and that weight loss maintenance may require the use of more strategies by some individuals than others.


Excitatory amino acid antagonists alleviate convulsive and toxic properties of lindane in mice.
Pesticides acting at GABAA receptors may induce convulsions in man and animals, but the mechanisms responsible for their convulsant activity are not fully explained. The following excitatory amino acid antagonists were studied for their protective action in mice intoxicated with chlorinated hydrocarbon insecticide lindane (gamma-hexachlorocyclohexane): the competitive NMDA antagonist: 3-(2-carboxypiperazine-4-yl)propenyl-1- phosphonic acid (D-CPPene, 20 mg/kg), the non-competitive NMDA antagonist: dizocilpine (MK-801, 0.4 mg/kg), the glycine site antagonist of NMDA receptor: 2-phenyl-1,3-propane-diol dicarbamate (felbamate, 400 mg/kg) and the competitive AMPA antagonist: 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX, 100 mg/kg). Systemic administration of an antagonist prior to lindane resulted in a strong anticonvulsant effect. D-CPPene, MK-801 and NBQX produced a marked increase of CD50 values of lindane for clonic convulsions. All the antagonists protected animals against tonic convulsions. Toxicity of lindane was potently reduced, as assessed 2, 24 and 120 hr after administration of the pesticide. Our results demonstrate that excitatory amino acid antagonists reduce convulsant properties and toxicity of lindane, suggesting that excitatory amino acid neurotransmission may be involved in its central action.


Ossification of the posterior longitudinal ligament in a Nigerian woman.
Ossification of the posterior longitudinal ligament (OPLL), though commonly reported among the Japanese population, has however been infrequently reported among other racial groups. It has not been reported among black Africans. A case of OPLL of the cervical spine seen in a 62-year-old Nigerian woman is hereby reported.


Programmed cell death promotes male sterility in the functional dioecious Opuntia stenopetala (Cactaceae).
The sexual separation in dioecious species has interested biologists for decades; however, the cellular mechanism leading to unisexuality has been poorly understood. In this study, the cellular changes that lead to male sterility in the functionally dioecious cactus, Opuntia stenopetala, are described. The spatial and temporal patterns of programmed cell death (PCD) were determined in the anthers of male and female flowers using scanning electron microscopy analysis and histological observations, focusing attention on the transition from bisexual to unisexual development. In addition, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assays were used as an indicator of DNA fragmentation to corroborate PCD. PCD was detected in anthers of both female and male flowers, but their patterns differed in time and space. Functionally male individuals developed viable pollen, and normal development involved PCD on each layer of the anther wall, which occurred progressively from the inner (tapetum) to the outer layer (epidermis). Conversely, functional female individuals aborted anthers by premature and displaced PCD. In anthers of female flowers, the first signs of PCD, such as a nucleus with irregular shape, fragmented and condensed chromatin, high vacuolization and condensed cytoplasm, occurred at the microspore mother cell stage. Later these features were observed simultaneously in all anther wall layers, connective tissue and filament. Neither pollen formation nor anther dehiscence was detected in female flowers of O. stenopetala due to total anther disruption. Temporal and spatial changes in the patterns of PCD are responsible for male sterility of female flowers in O. stenopetala. Male fertility requires the co-ordination of different events, which, when altered, can lead to male sterility and to functionally unisexual individuals. PCD could be a widespread mechanism in the determination of functionally dioecious species.


Lung ultrasound during hemodialysis: the role in the assessment of volume status.
Fluid balance is important in patients undergoing hemodialysis. "Dry" weight is usually estimated clinically, and also, bioimpedance is considered reliable. Ultrasonography of inferior vena cava (IVC) estimates central venous pressure, and lung ultrasound evaluates extravascular (counting B-lines artifact) lung water. Our study was aimed to clarify their usefulness in the assessment of volume status during hemodialysis. A total of 71 consecutive patients undergoing hemodialysis underwent lung and IVC ultrasound and bioimpedance spectroscopy immediately before and after dialysis. There was a significant reduction in the number of B-lines (3.13 vs 1.41) and in IVC diameters (end-expiratory diameter 1.71 vs 1.37; end-inspiratory diameter 1.19 vs 0.95) during dialysis. The reduction in B-lines correlated with weight reduction during dialysis (p 0.007); none of the parameters concerning the IVC correlated with fluid removal. At the end of the dialysis session, the total number of B-lines correlated with bioimpedance residual weight (p 0.002). The reduction in B-lines correlated with fluid loss due to hemodialysis, despite the small pre-dialysis number, confirming that lung ultrasound can identify even modest variations in extravascular lung water. IVC ultrasound, which reflects the intravascular filling grade, might not be sensitive enough to detect rapid volume decrease. Clinically estimated dry weight had a poor correlation with both bioimpedance and ultrasound techniques. Post-dialysis B-lines number correlates with residual weight assessed with bioimpedance, suggesting a role for ultrasound in managing hemodialysis patients.


Pulmonary pathology of patients treated with partial liquid ventilation.
Initial trials of partial liquid ventilation (PLV), which is gas ventilation of perfluorocarbon-filled lungs, are underway in patients with severe respiratory failure. We report the first study of the effects of the perfluorocarbon, perflubron, on the lung. Necropsies were conducted in nine patients (seven adults and two neonates; mean adult age, 31 +/- 5 yr) managed with PLV (average number of doses, 4 +/- 1). All of the patients required extracorporeal life support. The patients had pneumonia with the acute respiratory distress syndrome (six patients), trauma/capillary leak syndrome (one patient), congenital diaphragmatic hernia (one patient), and primary pulmonary hypertension (one patient). Nine adult patients (mean age, 37 +/- 5 yr) with acute respiratory distress syndrome requiring extracorporeal life support served as a control. Pathologic findings were evaluated in both groups. Lung weights in the adult patients of both groups were elevated (mean weight of PLV-treated right lung, 1401 +/- 186 g; mean weight of PLV-treated left lung, 1131 +/- 177 g; mean weight of control right lung, 1018 +/- 91 g; mean weight of control left lung, 988 +/- 80 g). There was no significant difference between the two groups (right lung, P = .066; left lung, P = .436). Frequent gross findings included focal consolidation, patchy hemorrhage, and glassy cut surfaces. The histologic findings were similar in both groups. Diffuse alveolar damage (either proliferative phase or mixed proliferative and exudative phases) was seen in all nine of the study patients. Eight of the nine control patients had diffuse alveolar damage (five had proliferative phase only, one had mixed proliferative and exudative phases, and two had exudative phase only). One other patient had extensive parenchymal necrosis. Other frequent findings were intra-alveolar hemorrhage, numerous intra-alveolar macrophages, and organization of exudate. PLV with perflubron in patients with adult and neonatal respiratory distress syndromes is not associated with unique pathologic findings in the human lung.


Clinical study of 10 cases of acute or subacute interstitial pneumonia associated with dermatomyositis.
Abstract The prognosis for dermatomyositis (DM) with acute interstitial pneumonia (IP) is very poor. In the past 5 years, we have treated 10 DM patients with acute or subacute IP. Six cases were of acute-type IP, and 4 were of subacute-type IP. The treatment was a combination therapy of methylprednisolone (m-PSL) pulse therapy, cyclophosphamide (CPA) pulse therapy, oral cyclosporine A (CsA), and oral PSL. The outcome was 5 deaths and 5 survivals. All 5 cases of death had acute-type IP, four of which were complicated with pneumomediastinum, and these patients died within 40 days of IP onset. Furthermore, 4 of the 5 death cases were diagnosed with amyopathic DM, and one had hypomyopathic DM. The survivors comprised one case of acute-type IP with marked myositis, and 4 subacute cases. These results suggested that the prognosis for DM with IP might be dependent on the type of IP, the severity of the myositis, and the existence of pneumomediastinum. The rapid establishment of a more useful diagnostic technique and therapy for early-phase DM with acute IP is hoped for.


A new United States Pharmacopeia (USP) Chapter 1046: cell and gene therapy products.
The United States Pharmacopeia (USP) has published the first draft of a new information chapter on cell and gene therapy products in January, 2000. The chapter discusses the manufacturing and testing cell and gene therapy products. It is intended for people working in the field as well as for pharmacists and clinicians who would like more information on this subject. This article outlines the background of USP and explains how it became involved with cell and gene therapy and what an information chapter is. It details the subjects covered by the chapter and the philosophy behind the chapter. This draft will be revised based on comments received from the public and then published as a revision subject to further comments before it becomes an official chapter in a supplement to USP 24-NF 19.


Factors affecting the emotional wellbeing of the caregivers of dementia sufferers.
Recent research on the factors that mediate the emotional wellbeing of the caregivers of dementia sufferers is reviewed. The roles of such factors as the caregiver's attributional style and coping strategies, the caregiver's relationship with the dementia sufferer, and levels of formal and informal support are discussed with reference to identifying those caregivers who are particularly vulnerable to emotional disorder or strain.


Lipophilic and respiratory properties of NADH and succinate dehydrogenase sites in mitochondria from various tissues of the rat.
Mitochondria were isolated from heart, liver, kidney, spleen, and brain of the rat. With overall regard to both resting and activated respiratory velocities with either glutamate or succinate, as well as the respective degrees of respiratory control, kidney mitochondria were most efficient and spleen mitochondria least so. A probe of mitochondrial inner membrane lipophilicity with tetrabutylammonium bromide showed that NADH dehydrogenases from liver and heart were similar, as were also those from kidney and spleen. With the exception of brain, only small differences were observed in lipophilic properties of succinate dehydrogenases from the various other tissues. Variation in lipophilic characteristics of the two sites on the mitochondrial inner membranes could not be correlated with embryological origin of the tissue.


FDG-PET reveals improved cardiac regeneration and attenuated adverse remodelling following Sitagliptin + G-CSF therapy after acute myocardial infarction.
Dual therapy comprising G-CSF for mobilization of bone marrow-derived progenitor cells (BMPCs), with simultaneous pharmacological inhibition of dipeptidylpeptidase-IV for enhanced myocardial recruitment of circulating BMPC via the SDF-1α/CXCR4-axis, has been shown to improve survival after acute myocardial infarction (AMI). Using an innovative method to provide non-invasive serial in vivo measurements and information on metabolic processes, we aimed to substantiate the possible effects of this therapeutic concept on cardiac remodelling after AMI using 2-deoxy-2-[18F]fluoro-d-glucose positron emission tomography (FDG-PET). AMI was induced in C57BL/6 mice by performing surgical ligation of the left anterior descending artery in these mice. Animals were then treated with granulocyte-colony stimulating factor + Sitagliptin (GS) or placebo for a duration of 5 days following AMI. From serial PET scans, we verified that the infarct size in GS-treated mice (n = 13) was significantly reduced at Day 30 after AMI when compared with the mice receiving placebo (n = 10). Analyses showed a normalized FDG uptake on Day 6 in GS-treated mice, indicating an attenuation of the cardiac inflammatory response to AMI in treated animals. Furthermore, flow cytometry showed a significant increase in the anti-inflammatory M2-macrophages subpopulation in GS-treated animals. In comparing GS treated with placebo animals, those receiving GS-therapy showed a reduction in myocardial hypertrophy and left ventricular dilatation, which indicates the beneficial effect of GS treatment on cardiac remodelling. Remarkably, flow cytometry and immunohistochemistry showed an increase of myocardial c-kit positive cells in treated mice (n = 12 in both groups). Using the innovative method of micro-PET for non-invasive serial in vivo measurements of metabolic myocardial processes in mice, we were able to provide mechanistic evidence that GS therapy improves cardiac regeneration and reduces adverse remodelling after AMI.


The Effect of a Short-Term Exposure to Lead on the Levels of Essential Metal Ions, Selected Proteins Related to Them, and Oxidative Stress Parameters in Humans.
The present study was designed to explore the possible influence of subacute exposure to lead on the levels of selected essential metals, selected proteins related to them, and oxidative stress parameters in occupationally exposed workers. The study population included 36 males occupationally exposed to lead for 36 to 44 days. Their blood lead level at the beginning of the study was 10.7 ± 7.67 μg/dl and increased to the level of 49.1 ± 14.1 μg/dl at the end of the study. The levels of calcium, magnesium, and zinc increased significantly after lead exposure compared to baseline by 3%, 3%, and 8%, respectively, while the level of copper decreased significantly by 7%. The malondialdehyde (MDA) level and the activities of catalase (CAT) and superoxide dismutase (SOD) did not change due to lead exposure. However, the level of lipid hydroperoxides (LPH) in serum increased significantly by 46%, while the level of erythrocyte lipofuscin (LPS) decreased by 13%. The serum levels of essential metals are modified by a short-term exposure to lead in occupationally exposed workers. A short-term exposure to lead induces oxidative stress associated with elevated levels of LPH but not MDA.


[Towards eradication of poliomyelitis in Vietnam and the western Pacific region, 1993].
In 1988, the World Health Assembly established the goal of global eradication of poliomyelitis by the year 2000, and the Western Pacific Region (WPR) of WHO established the goal of regional eradication by 1995. Countries in the WPR with endemic poliomyelitis include China, Viet Nam, Cambodia, Laos, and the Philippines. Each country has a national plan of action for eradication, based on three main strategies: maintenance of high coverage with three doses of OPV through routine immunization; supplemental immunization with OPV, including annual National Immunization Days (NIDs); and surveillance of acute flaccid paralysis cases, including isolation of polioviruses in stool specimens. The number of confirmed poliomyelitis cases reported to WHO decreased from 2,635 cases in 1991 to 1,226 cases in 1993, mostly reflecting the decrease in China where supplemental immunization started in 1990 in high-incidence provinces. In six provinces in Viet Nam where immunization days took place in 1992, confirmed cases dropped from 168 in 1992 to 42 in 1993. China, the Philippines, Viet Nam and Laos have conducted NIDs in 1993-1994, with two doses of OPV administered to more than 100 millions children under 5 years of age. NIDs will be repeated in 1994 and 1995, and are planned in Cambodia in January-February 1995. NIDs receive strong support from national governments, and financial assistance from Rotary International, Unicef, the governments of Australia, Japan and Canada, and the US CDC. The apparent elimination of wild polioviruses from the Americas, and the substantial progress already made in the WPR, demonstrate the feasibility of eradicating polio in the WPR and other regions of the world.


[Consequences of stimulation on oocyte quality].
In all IVF programs, ovarian stimulation may lead to a hyperstimulation syndrome. Mild ovarian stimulations are suggested to reduce this complication of infertility treatment. In this article, we wonder what the consequences of different stimulations are on oocytes quality. The quality of the oocyte being the major factor of the development of a good embryo, it can be evaluated by direct or indirect techniques. Some are subjective. Several studies seem to show that ovarian stimulation has an influence on oocye quality, especially at a chromosomic and perharps, epigenetic level. It seems that a mild stimulation (for example with GnRH antagonist) selects less oocytes but with a better quality and produces euploid embryos. The uterine transfer of this kind of embryo should lead to an ongoing pregnancy.


Dextromethorphan for neuropathic pain with Churg-Strauss syndrome.
Abstract A 48-year old man who had been treated with prednisolone, aminophylline, and pranlukast, a leukotriene-receptor antagonist, was diagnosed with Churg-Strauss syndrome based on the findings of asthma, eosinophilia, mononeuropathy, and extravascular eosinophils. Intractable neuropathic pain of the legs was successfully controlled with an N-methyl-d-aspartate receptor antagonist, dextromethorphan. We suggest that dextromethorphan receptor antagonists may be a useful treatment for the pain of neuropathy caused by vasculitis syndrome.


Comparative Genomic Analysis Provides Insights into the Phylogeny, Resistome, Virulome, and Host Adaptation in the Genus Ewingella.
Ewingella americana is a cosmopolitan bacterial pathogen that has been isolated from many hosts. Here, we sequenced a high-quality genome of E. americana B6-1 isolated from Flammulina filiformis, an important cultivated mushroom, performed a comparative genomic analysis with four other E. americana strains from various origins, and tested the susceptibility of B6-1 to antibiotics. The genome size, predicted genes, and GC (guanine-cytosine) content of B6-1 was 4.67 Mb, 4301, and 53.80%, respectively. The origin of the strains did not significantly affect the phylogeny, but mobile genetic elements shaped the evolution of the genus Ewingella. The strains encoded a set of common genes for type secretion, virulence effectors, CAZymes, and toxins required for pathogenicity in all hosts. They also had antibiotic resistance, pigments to suppress or evade host defense responses, as well as genes for adaptation to different environmental conditions, including temperature, oxidation, and nutrients. These findings provide a better understanding of the virulence, antibiotic resistance, and host adaptation strategies of Ewingella, and they also contribute to the development of effective control strategies.


Bayesian approach to assay sensitivity analysis of thorough QT trials.
One of the analyses recommended in ICH E14 Guidance (International Conference on Harmonisation, 2005) after the test drug is shown to be negative in QT interval prolongation after subjects treated with the test drug is an assay sensitivity analysis of a positive control drug with known effect on QT prolongation. The assay sensitivity is validated if the response profile is shown to be expected and the QT interval after administration of the positive control drug is shown to be at least 5 ms more than placebo. The negative result of the test treatment is validated if the prolongation of the positive control is verified among the study subjects. One of the most frequently used positive control drugs in thorough QT (TQT) trials is moxifloxacin. In order to improve the efficiency of the study and to reduce the number of subjects exposed to moxifloxacin, we explore the potential sample size reduction with a Bayesian approach to the assay sensitivity utilizing the data of historical TQT trials. We derived the distribution of moxifloxacin-induced QT prolongation based on 14 crossover trials and six parallel trials. The estimated distribution is used as a prior distribution to assess the posterior probability that the moxifloxacin-induced QT prolongation is larger than 5 ms. Sample size based on such Bayesian approach will be compared with the conventional frequentist approach for efficiency assessment.


FIXATION PROBABILITIES OF SELFING RATE MODIFIERS IN SIMULATIONS WITH SEVERAL DELETERIOUS ALLELES WITH LINKAGE.
The fixation rates of selfing rate modifiers were found by stochastic simulation in an infinite site model, including effects of several deleterious alleles with variable effects, which were randomly distributed in the genome without assuming any pollen discounting. Previous results on the evolution of selfing obtained by more precise methods were in this study further validated, and it was concluded that the effect of genetic associations on the evolution of mating systems is small except in the case of full pollen discounting. Furthermore, attention was given to the uneven distribution of the genetic load in the population, and the accompanying large among-genome variation in fixation rates. This among-genome variation will be of significance for the evolution of mating systems.


Automatic infection detection system.
An infected person may be contagious already before the first symptoms appear. This person can, in the period of disease evolution, infect several associated citizens before consulting a general practitioner (GP). Early detection of contagion is therefore important to prevent spreading of diseases. The Automatic Infection Detection (AID) System faces this problem through investigating the hypothesis that the blood glucose (BG) level increases when a person is infected. The first objective of the prototyped version of the AID system was to identify possible BG elevations in the incubation time that could be related to the spread of infectious diseases. To do this, we monitored two groups of people, with and without diabetes mellitus. The AID system analyzed the results and we were able to detect two cases of infection during the study period. The time of detection occurred simultaneous or near the time of onset of symptoms. The detection did not occur earlier for a number of reasons. The most likely one is that the evolution process of an infectious disease is both complicated and involves the immune system and several organs in the body. The investigation with regard to isolating the key relations is therefore considered as a very complex study. Nevertheless, the AID system managed to detect the infection much earlier than what is possible with today's early warning systems for infectious diseases.


Gold nanoparticle-embedded porous graphene thin films fabricated via layer-by-layer self-assembly and subsequent thermal annealing for electrochemical sensing.
A uniform three-dimensional (3D) gold nanoparticle (AuNP)-embedded porous graphene (AuEPG) thin film has been fabricated by electrostatic layer-by-layer assembly of AuNPs and graphene nanosheets functionalized with bovine serum albumin and subsequent thermal annealing in air at 340 °C for 2 h. Scanning electron microscopy (SEM) investigations for the AuEPG film indicate that an AuNP was embedded in every pore of the porous graphene film, something that was difficult to achieve with previously reported methods. The mechanism of formation of the AuEPG film was initially explored. Application of the AuEPG film in electrochemical sensing was further demonstrated by use of H(2)O(2) as a model analyte. The AuEPG film-modified electrode showed improved electrochemical performance in H(2)O(2) detection compared with nonporous graphene-AuNP composite film-modified electrodes, which is mainly attributed to the porous structure of the AuEPG film. This work opens up a new and facile way for direct preparation of metal or metal oxide nanoparticle-embedded porous graphene composite films, which will enable exciting opportunities in highly sensitive electrochemical sensors and other advanced applications based on graphene-metal composites.


[Gene-gene interaction on central obesity in school-aged children in China].
Objective: To investigate possible effect of 6 obesity-associated SNPs in contribution to central obesity and examine whether there is an interaction in the 6 SNPs in the cause of central obesity in school-aged children in China. Methods: A total of 3 502 school-aged children who were included in Beijing Child and Adolescent Metabolic Syndrome (BCAMS) Study were selected, and based on the age and sex specific waist circumference (WC) standards in the BCAMS study, 1 196 central obese cases and 2 306 controls were identified. Genomic DNA was extracted from peripheral blood white cells using the salt fractionation method. A total of 6 single nucleotide polymorphisms (FTO rs9939609, MC4R rs17782313, BDNF rs6265, PCSK1 rs6235, SH2B1 rs4788102, and CSK rs1378942) were genotyped by TaqMan allelic discrimination assays with the GeneAmp 7900 sequence detection system (Applied Biosystems, Foster City, CA, USA). Logistic regression model was used to investigate the association between 6 SNPs and central obesity. Gene-gene interactions among 6 polymorphic loci were analyzed by using the Generalized Multifactor Dimensionality Reduction (GMDR) method, and then logistic regression model was constructed to confirm the best combination of loci identified in the GMDR. Results: After adjusting gender, age, Tanner stage, physical activity and family history of obesity, the FTO rs9939609-A, MC4R rs17782313-C and BDNF rs6265-G alleles were associated with central obesity under additive genetic model (OR=1.24, 95%CI: 1.06-1.45, P=0.008; OR=1.26, 95%CI: 1.11-1.43, P=2.98×10(-4); OR=1.18, 95% CI: 1.06-1.32, P=0.003). GMDR analysis showed a significant gene-gene interaction between MC4R rs17782313 and BDNF rs6265 (P=0.001). The best two-locus combination showed the cross-validation consistency of 10/10 and testing accuracy of 0.539. This interaction showed the maximum consistency and minimum prediction error among all gene-gene interaction models evaluated. Moreover, the combination of MC4R rs17782313-C and BDNF rs6265-G was associated with an increased risk of central obesity after adjustment for gender, age, Tanner stage, physical activity and family history of obesity. Conclusions: Our study showed that FTO rs9939609-A, MC4R rs17782313-C and BDNF rs6265-G alleles were associated with central obesity, and statistical interaction between MC4R rs17782313-C and BDNF rs6265-G increased risk of central obesity in school-aged children in China.


Development of a novel NS1 competitive enzyme-linked immunosorbent assay for the early detection of Zika virus infection.
Zika virus (ZIKV) is a flavivirus that has emerged as a global health threat after the 2015 outbreak in the Americas, where devastating congenital defects were documented. There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to identify ZIKV without cross-reactive interference of related flaviviruses. Early diagnosis is critical when treating symptomatic patients and in preventing ZIKV transmission. In this context, the development of sensitive and accurate diagnostic methods are urgently needed for the detection of ZIKV acute infection. The aim of this study consisted of obtaining monoclonal antibodies (mAbs) against denatured monomeric ZIKV Nonstructural protein 1 (ZNS1), a useful diagnostic marker for flavivirus early detection, in order to develop a highly specific and sensitive ZNS1 indirect competitive ELISA (icELISA). The production of hybridomas secreting ZNS1 mAbs was carried out through immunizations with denatured monomeric ZNS1. We selected 1F5 and 6E2 hybridoma clones, which recognized the heat-denatured ZNS1 hexameric form by indirect ELISA. Cross-reaction studies indicated that these mAbs specifically bind to a ZNS1 linear epitope, and that they do not cross-react with the NS1 protein from other related flaviviruses. The 1F5 mAb enabled the development of a sensitive and reproducible icELISA to detect and quantify small amounts of ZNS1 disease marker in heat-denatured human sera. Here, we establish a reliable 1F5 based-icELISA that constitutes a promising diagnostic tool for control strategies and the prevention of ZIKV propagation.


Cardiac catheterization and operative outcomes from a multicenter consortium for children with williams syndrome.
Williams syndrome is a multifaceted disorder that includes a spectrum of cardiovascular anomalies. Due to its rare occurrence, outcome data for operations and cardiac catheterization are limited. We reviewed data from 242 individuals from the Pediatric Cardiac Care Consortium (PCCC) with Williams syndrome and associated cardiovascular lesions, and their frequency, and assessed their effects on mortality. In the PCCC, from 1984 to 1999 there were approximately 100,000 entries for cardiac procedures, involving more than 62,000 patients. The diagnosis of Williams syndrome was based on clinical features and determined by each site. Most patients were diagnosed with the availability of the FISH probe for region 7q11.23. Using a spreadsheet application, Microsoft Excel, the selected patients were analyzed for various types of cardiac anomalies. The most common cardiovascular lesions and the mortality rate in patients with Williams syndrome were examined. A complete tabulation of all cardiovascular lesions was assembled. There were 292 catheterizations and 143 operations reported to the PCCC. One hundred six patients had both an operation and a catheterization. The three main cardiovascular anomalies were supravalvular aortic stenosis (SVAS; 169), pulmonary artery stenosis (PAS; 130), and coarctation or aortic arch hypoplasia (Arch; 32). One hundred five patients had a single lesion, 70 with SVAS, 29 with PAS, and 6 with an arch anomaly. Ninety-two had more than one lesion: 80 with SVAS and PAS, 7 with PAS and Arch, and 5 with SVAS and Arch. Seventy individuals have only SVAS, 29 PAS, and 6 Arch alone. There was a total of 15 deaths. The mortality rate was highest in the group with the combination of SVAS and PAS (7 surgical and 5 catheter; 12 of 80 patients [15%]; p = 0.0001, chi(2)). In conclusion, our data represent the largest collection of individuals with Williams syndrome who underwent cardiac catheterization and/or operation. The data suggest that children with Williams syndrome and bilateral outflow tract obstruction have statistically and clinically significantly higher mortality associated with catheterization or operation.


PCNA in the ovary of zebrafish (Brachydanio rerio, Ham.-Buch.).
PCNA, a nuclear protein intimately involved in DNA synthesis, has been localized immunohistochemically in ovaries of zebrafish (Brachydanio rerio, Ham.-Buch.). Positively stained cells were detected in the follicle epithelium of all stages of growing oocytes and degenerating oocytes. Furthermore, PCNA was found to be present in nuclei of oogonia and oocytes in meiotic prophase. Follicle epithelium showed an increasing number of PCNA-positive cells during egg cell development with a very low index of <0.5% at the beginning and a maximum of >7% in stage IV oocytes that are rapidly incorporating yolk protein. In this phase, follicle epithelium often showed groups of stained cells in limited zones of proliferation. In degenerating oocytes, over 60% of the former follicle cells were positive for PCNA. Nuclei of oogonia were always positive and the nucleus of stage III oocytes showed strong positivity. Stage III is the stage in which amplification of rRNA genes occurs and numerous nucleoli apear in the peripheral caryoplasm. It is concluded that PCNA expression is related to amplification of rDNA.


Novel bioactive oxazolomycin isomers produced by Streptomyces albus JA3453.
Two novel oxazolomycin isomers, oxazolomycins B (2) and C (3), were isolated from the fermentation broth of an oxazolomycin-producing strain, Streptomyces albus JA3453. Both compounds are geometrical isomers of oxazolomycin (1), the configurations of their triene moieties being (4'E, 6'E, 8'E) (2) and (4'Z, 6'E, 8'E) (3) while that of oxazolomycin (1) is (4'Z, 6'Z, 8'E). Compounds 2 and 3 exhibited potent inhibitory activity against crown gall formation with the same MIC (0.8 microgram/disk) as oxazolomycin. Compounds 2 and 3 showed no antibacterial activity against Agrobacterium tumefaciens, in contrast to oxazolomycin which has specific anti-A. tumefaciens activity.


Extra-amniotic saline infusion increases cesarean risk versus other induction methods and spontaneous labor.
Extra-amniotic saline infusion (EASI) via a Foley catheter has been thought to be comparable in efficacy to other induction and cervical ripening methods. This study examines the risk of cesarean delivery associated with EASI compared with spontaneous labor and other methods of cervical ripening. A retrospective cohort study based upon deliveries at Bellevue Hospital Center from August 2000 to December 2002 was conducted. Three groups were identified: EASI, other methods of induction such as prostaglandins and oxytocin administration, and spontaneous labor. Pairwise comparisons were performed using analysis of variance and multivariate logistic regression analysis to control for confounding variables. There were 625 charts evaluated: including 171 with EASI, 190 with other induction methods, and 264 with spontaneous labor. The rates of cesarean section were 33.9%, 17.9%, and 7.2%, respectively. When compared with spontaneous labor, there was a higher risk of cesarean delivery for subjects induced with other methods (adjusted odds ratio [OR], 2.4; 95% confidence interval [CI], 1.3 to 4.5; P < 0.001), and for those induced with EASI (adjusted OR, 5.5; 95% CI, 3.1 to 9.9; P < 0.001). When EASI was compared with other methods of induction, the risk of cesarean delivery was still increased (adjusted OR, 2.3; 95% CI, 1.4 to 3.8; P = 0.001). EASI is associated with an increased risk of cesarean delivery compared with spontaneous labor and other methods of cervical ripening.


Early detection of sequela-prone osteomyelitis in children with use of simple clinical and laboratory criteria.
To determine which clinical or laboratory criteria best reflected the prognosis for 83 children with acute hematogenous osteomyelitis (AHO), they were compared with outcomes after a follow-up of at least 2 months (for 78%, > or = 6 months). Twenty-eight children (34%) developed sequelae. They had higher serum C-reactive protein (CRP) concentrations (days 1-6 of treatment; P = .0004 to .0001) and higher clinical scores (P = .0001) than did patients who had an uneventful recovery. The frequency of sequelae increased from 3% to 73% (P = .0001) when CRP concentrations exceeded the defined cutoff limits and the clinical scores were > or = 1. Age, the duration of symptoms at diagnosis, and the type and duration of intravenous antimicrobial therapy or surgical management did not differ (P > .05) between children with and without sequelae. Both CRP determinations and clinical evaluations with use of a scoring system enable early detection of sequela-prone AHO in children and are most accurate when used together.


Effects of drugs on the pyrogenicity of toxic shock syndrome toxin 1 and its capacity to enhance susceptibility to the lethal effects of endotoxic shock in rabbits.
The effects of various kinds of drugs on the pyrogenicity of toxic shock syndrome toxin 1 (TSST-1) and its capacity to enhance the lethal effects of endotoxin in rabbits were studied. Antipyretic agents--including aspirin, aminopyrine, and indomethacin--that were given immediately or 2 hours after injection of TSST-1 decreased the febrile response to TSST-1 but did not inhibit the enhancement of lethality. Cyclosporine, which was given 2 hours after injection of TSST-1, suppressed the fever but did not inhibit the lethal effects. Methylprednisolone given immediately or 2 hours after TSST-1 injection did not decrease the fever but inhibited the lethal effect. Of the agents phentolamine, dopamine, and chlorpromazine, which are reported to suppress endotoxic shock experimentally, only chlorpromazine decreased the fever, and none inhibited the lethal effects.