Effect of tetrahydrobiopterin on selective endothelial dysfunction of epicardial porcine coronary arteries induced by cardiopulmonary bypass.
We hypothesized that cardiopulmonary bypass induces a selective alteration of the coronary arterial endothelial cell signal transduction which could be explained by a state of depletion and/or decreased activity of endogenous tetrahydrobiopterin (BH(4)). The aim of this study was to assess the effects of cardiopulmonary bypass and BH(4) on the endothelial function of epicardial coronary arteries in a swine model of cardiopulmonary bypass. Swine underwent 90 min of cardiopulmonary bypass alone (N=19) or in association with a brief cardioplegic arrest with (N = 6) or without (N = 5) in vivo BH(4) administration, followed by a 60-min period following weaning from cardiopulmonary bypass and were compared to a control group (N = 7). Endothelium-dependent relaxations of epicardial coronary artery rings were studied using standard organ chamber experiments in the presence or absence of in vitro BH(4) or superoxide dismutase (SOD) and catalase. Cardiopulmonary bypass caused a statistically significant reduction of endothelium-dependent relaxations to serotonin (p < 0.0001), bradykinin (p < 0.001), UK14304 (p < 0.0001) and calcium ionophore (p < 0.01) in epicardial porcine coronary arteries. In vitro and in vivo BH(4) supplementation improved endothelium-dependent relaxations to serotonin and bradykinin, which were left unchanged by SOD-catalase administration. Cardiopulmonary bypass was associated with a decrease in nitric oxide availability (p = 0.002) and increased oxidative stress (p < 0.001), which were both restored by in vivo BH(4) administration (p < 0.001). Treatment with BH(4) improves the endothelial dysfunction of porcine epicardial coronary arteries, restores nitric oxide availability and reduces the oxidative stress associated with cardiopulmonary bypass.


Biosorption of nickel onto treated alga (Oedogonium hatei): Application of isotherm and kinetic models.
Oedogonium hatei was developed into an effective and efficient adsorbent for the removal of Ni(II) ions from aqueous solution. The adsorption studies of untreated and treated algal biomass (with 0.1M HCl) were compared in batch mode. Optimum biosorption conditions were determined as a function of pH, biomass dosage, contact time, and temperature and the Langmuir and Freundlich isotherms were found applicable in terms of relatively high regression values. The maximum monolayer adsorption capacity of the biosorbents (untreated and acid-treated algae), as obtained from the Langmuir adsorption isotherm, was found to be 40.9 and 44.2mg/g, respectively at 80min contact time, 5.0 pH, 0.7g/L algal dose, and 298K temperature. The thermodynamic parameters showed that the adsorption of Ni(II) ions onto algal biomass was feasible, spontaneous, and exothermic under the studied conditions. Kinetics of adsorption followed both first- and second-order rate equations and the process involving the rate-controlling step is complex involving boundary layer as well as intraparticle diffusion processes. The FTIR results of algal biomass showed that biomass has different functional groups and these functional groups are able to react with metal ion in aqueous solution. Biosorbent could be regenerated using 0.1M NaOH solution, with up to 70% recovery. The performance of this biosorbent was then compared with many other reported biosorbents for nickel removal and it was observed that the proposed adsorbent is effective in terms of its performance.


A comparative analysis of the cycle fecundity rates associated with combined human menopausal gonadotropin (hMG) and intrauterine insemination (IUI) versus either hMG or IUI alone.
Human menopausal gonadotropin (hMG) superovulation combined with washed intrauterine insemination (IUI) has been advocated for the treatment of various forms of infertility when more traditional therapy has failed. To assess the relative efficacy of combined treatment with hMG and IUI compared with either hMG or IUI alone, pregnancy outcomes of the three treatment groups were compared in couples having infertility because of male factor, cervical factor, endometriosis, or unexplained. A total of 751 cycles were analyzed from 322 couples. The mean cycle fecundity rate associated with hMG/IUI therapy was significantly higher than either hMG or IUI therapy alone for all patients (hMG/IUI = 19.6%, hMG = 6.3%, IUI = 3.4%). The improvement in cycle fecundity rates with hMG/IUI therapy was also observed when the couples were separated by infertility diagnostic groups: male factor (hMG/IUI = 15.3%, hMG = 4.4%, IUI = 3.0%), cervical factor (hMG/IUI = 26.3%, hMG = 7.9%, IUI = 5.1%), endometriosis (hMG/IUI = 12.85%, hMG = 6.6%), and unexplained infertility (hMG/IUI = 32.6%, hMG = 5.5%, IUI = 0%). Moreover, in patients who had failed to conceive with hMG or IUI alone, the cycle fecundity rate when they were switched to hMG/IUI therapy equaled that of patients who received combined therapy from the onset. We conclude that cycle fecundity rates and cumulative pregnancy rates are significantly greater using a combination of hMG and IUI compared with either modality alone in the treatment of male factor, cervical factor, endometriosis, or unexplained infertility. Indeed, in couples with nontubal related infertility, cycle fecundity rates with hMG/IUI approach the rates seen with in vitro fertilization and gamete intrafallopian tube transfer.


Isolation, Culture, and Staining of Single Myofibers.
Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation.


Valproic acid and thrombocytopenia: cross-sectional study.
To investigate the relationship between platelet count and serum valproic acid level, age, duration of valproic acid therapy, and polytherapy, and to determine the clinical significance of thrombocytopenia associated with high-dosage valproic acid therapy. Cross-sectional study. Residential unit for neurologically impaired children and paediatric out-patient clinic, Hong Kong. Ninety-six neurologically impaired children who were treated with valproic acid between 1 July 1991 to 3 June 1999. The comparison group consisted of 48 children receiving antiepileptic drugs other than valproic acid. Low- or high-dosage valproic acid, using the threshold value of 40 mg/kg/d. Platelet count and liver function, duration of valproic acid treatment, dosage, and trough serum valproic acid concentration. Seventeen (17.7%) patients in the treatment group developed thrombocytopenia, compared with two (4.2%) in the comparison group (P<0.05). The platelet count was negatively correlated to serum valproic acid level and age, and positively correlated to polytherapy. The duration of valproic acid treatment was not a confounding factor in the age-related decrease in platelet count. Children with a trough level of >450 micromol/L or a daily dose of >40 mg/kg were more likely to develop thrombocytopenia. Thrombocytopenia was mild in most cases. A trough valproic acid level of >450 micromol/L or a daily dose of >40 mg/kg should alert the clinician to the risk of developing thrombocytopenia. The risk is further increased for older children. The platelet count should be monitored for patients receiving a high concentration of valproic acid who are also receiving drugs that would affect homeostasis, or who are undergoing surgical procedures.


[News in pathophysiology and management of preterm labour].
To summarize available data concerning pathophysiology and management of preterm labour and their use in clinical practice. Department of Obstetrics and Gynecology od the First Faculty of Medicine, Charles University, and General Teaching Hospital, Prague. Review article. Compilation od published data from scientific literature. Birth canal infections seem to play a key role in the ethiopathogenesis of premature delivery; the related biochemical changes significantly affect perinatal morbidity and mortality. Other potential causes, particularly hormone metabolism disorders or uteroplacental ischaemia have been intensively studied. The particular pathogenetic process of premature delivery is inflammation. This process is related to both mother and fetus. Fetal inflammatory response (FIRS)--can occur without maternal response--is connected with significant increase in perinatal morbidity. FIRS is characterised by defined laboratory, histological and clinical criteria. Effective primary prevention of premature delivery does not exist at present. The sensitivity and specificity of the laboratory markers having been used so far is low. Thus, the research is focused on finding new inflammation markers to allow early identification of pregnant women at a high risk of premature delivery and fetal inflammation. The screening of women at a high risk by means of new laboratory and ultrasound tests belongs to the most important steps in secondary and tertiary prevention of premature delivery. Intensive research of potential trigger mechanisms has been carried out, including a variety of gene types, which are potentially related to the process of premature delivery. We try to exercise new informations about pathophysiology of preterm labour in prenatal care, firstly about thrombophillias and gestagens. We also use a principles of "evidence based medicine" and revalue importance of steroids, tocolytics and antibiotics.


Integration of calcium and cyclic AMP signaling pathways by 14-3-3.
Calcium-stimulated nuclear factor of activated T cells (NFAT) transcription activity at the interleukin-2 promoter is negatively regulated by cyclic AMP (cAMP). This effect of cAMP is mediated, in part, by protein kinase A phosphorylation of NFAT. The mechanism of regulation involves the creation of a phosphorylation-dependent binding site for 14-3-3. Decreased NFAT phosphorylation caused by the calcium-stimulated phosphatase calcineurin, or mutation of the PKA phosphorylation sites, disrupted 14-3-3 binding and increased NFAT transcription activity. In contrast, NFAT phosphorylation caused by cAMP increased 14-3-3 binding and reduced NFAT transcription activity. The regulated interaction between NFAT and 14-3-3 provides a mechanism for the integration of calcium and cAMP signaling pathways.


Aspects of pharmacodynamics and biotransformation of the glucocorticoid resocortol butyrate.
Aspects of the biotransformation and pharmacodynamics of the novel glucocorticoid resocortol butyrate (RCB) and its metabolites were assessed in vitro and in vivo in comparison with selected reference compounds. The main route of biotransformation of ((3)H)-RCB in the skin and the liver was 5alpha-reduction of the A-ring followed by reduction of the 3-carbonyl group. In the liver, metabolism was much more rapid than in the skin and 5beta-reduction also occurred. RCB had a relative binding affinity for the glucocorticoid receptor similar to that of triamcinolone acetonide, about 1.5 times that of dexamethasone, three times that of betamethasone valerate (BMV) and 10-14 times that of cortisol. The metabolites of RCB displayed only low to very low affinities for the receptor. The suppression of the hypothalamic-pituitary-adrenal axis was investigated in placebo- and positive-controlled studies in dogs by measurement of basal and corticotrophin-releasing hormone (CRH) stimulated plasma cortisol concentrations. The AUC of the plasma cortisol vs. time curve following CRH stimulation, a measure of adrenal suppression, was reduced significantly after topical application of BMV compared with the pretreatment values. The AUC in the RCB group was not reduced significantly. Adrenocorticotrophic hormone concentrations were not affected. Oral administration of RCB did not suppress adrenocortical function, whereas BMV induced almost complete suppression of basal and CRH-induced cortisol concentrations. The pharmacodynamics of RCB makes it a relatively safe glucocorticosteroid for topical application.


Cholesterol modulates the membrane binding and intracellular distribution of annexin 6.
Annexins are Ca(2+)- and phospholipid-binding proteins that are widely expressed in mammalian tissues and that bind to different cellular membranes. In recent years its role in membrane traffic has emerged as one of its predominant functions, but the regulation of its intracellular distribution still remains unclear. We demonstrated that annexin 6 translocates to the late endocytic compartment in low density lipoprotein-loaded CHO cells. This prompted us to investigate whether cholesterol, one of the major constituents of low density lipoprotein, could influence the membrane binding affinity and intracellular distribution of annexin 6. Treatment of crude membranes or early and late endosomal fractions with digitonin, a cholesterol-sequestering agent, displayed a strong reduction in the binding affinity of a novel EDTA-resistant and cholesterol-sensitive pool of annexin 6 proteins. In addition, U18666A-induced accumulation of cholesterol in the late endosomal compartment resulted in a significant increase of annexin 6 in these vesicles in vivo. This translocation/recruitment correlates with an increased membrane binding affinity of GST-annexin 6 to late endosomes of U18666A-treated cells in vitro. In conclusion, the present study shows that changes in the intracellular distribution and concentration of cholesterol in different subcellular compartments participate in the reorganization of intracellular pools of Ca(2+)-dependent and -independent annexin 6.


High prevalence of lipopolysaccharide mutants and R2-pyocin susceptible variants in Pseudomonas aeruginosa populations sourced from cystic fibrosis lung infections.
Cystic fibrosis (CF) patients often experience chronic, debilitating lung infections caused by antibiotic-resistant Pseudomonas aeruginosa, contributing to antimicrobial resistance (AMR). The genetic and phenotypic diversity of P. aeruginosa populations in CF lungs raises questions about their susceptibility to non-traditional antimicrobials, like bacteriocins. In this study, we focused on R-pyocins, a type of bacteriocin with high potency and a narrow killing spectrum. Our findings indicate that a large number of infectious CF variants are susceptible to R2-pyocins, even within diverse bacterial populations, supporting their potential use as therapeutic agents. The absence of a clear correlation between lipopolysaccharide (LPS) phenotypes and R-pyocin susceptibility suggests that LPS packing density may play a significant role in R-pyocin susceptibility among CF variants. Understanding the relationship between LPS phenotypes and R-pyocin susceptibility is crucial for developing effective treatments for these chronic infections.


Antioxidative potential of Duranta repens (Linn.) fruits against H2O2 induced cell death in vitro.
The effects of Duranta repens fruits were investigated on H2O2 induced oxidative cell death to evaluate its antioxidative potential in vitro. HEK293T cells were treated with different concentrations [0-1000 µg/ ml] of ethanol extract (E-Ex) and methanol extract (M-Ex) of D. repens for 24h, and then treated with 100 µM H2O2 for 24h. Cell viability, antioxidant parameters of cells, and antioxidant constituents of the extracts were determined. Treatment with limited dose of E-Ex or M-Ex increased the survival rate of H2O2-treated HEK293T cells, however the extra-high dose showed growth inhibitory effect. Treatment with E-Ex or M-Ex protected cellular lipid per-oxidation. In vitro analyses showed the 2,2-diphenyl-1-picrylhydrazyl and H2O2 scavenging activities as well as reducing potential of the extracts. We report here that the limited dose of E-Ex and M-Ex possess antioxidative potential, which can protect H2O2-induced oxidative cell damage.


Isoquinoline alkaloids and their binding with DNA: calorimetry and thermal analysis applications.
Alkaloids are a group of natural products with unmatched chemical diversity and biological relevance forming potential quality pools in drug screening. The molecular aspects of their interaction with many cellular macromolecules like DNA, RNA and proteins are being currently investigated in order to evolve the structure activity relationship. Isoquinolines constitute an important group of alkaloids. They have extensive utility in cancer therapy and a large volume of data is now emerging in the literature on their mode, mechanism and specificity of binding to DNA. Thermodynamic characterization of the binding of these alkaloids to DNA may offer key insights into the molecular aspects that drive complex formation and these data can provide valuable information about the balance of driving forces. Various thermal techniques have been conveniently used for this purpose and modern calorimetric instrumentation provides direct and quick estimation of thermodynamic parameters. Thermal melting studies and calorimetric techniques like isothermal titration calorimetry and differential scanning calorimetry have further advanced the field by providing authentic, reliable and sensitive data on various aspects of temperature dependent structural analysis of the interaction. In this review we present the application of various thermal techniques, viz. isothermal titration calorimetry, differential scanning calorimetry and optical melting studies in the characterization of drug-DNA interactions with particular emphasis on isoquinoline alkaloid-DNA interaction.


Gonadal steroid hormone regulation of human and mouse follicle stimulating hormone beta-subunit gene expression in vivo.
Transgenic mice that express a 10-kilobase human FSH beta (hFSH beta) gene exclusively in pituitary gonadotropes were used to study the regulation of hFSH beta gene expression by gonadal steroids. For comparison, the mouse FSH beta (mFSH beta) gene was studied in parallel in nontransgenic sibling (normal) mice. The hFSH beta gene showed a sexually dimorphic expression pattern, identical to mFSH beta, in the mouse environment. Intact normal and transgenic male mice had elevated (P < 0.05) levels of serum [16 +/- 2 ng/ml (normal); 38 +/- 6 (transgenic)] and pituitary FSH content [2 +/- 0.3 micrograms/mg protein (normal); 36 +/- 6 (transgenic)] and FSH beta mRNA [1.47 +/- 0.10 arbitrary density units (normal); 1.00 +/- 0.23 (transgenic)] compared to the corresponding female mice ([< 2.0 ng/ml (normal and transgenic)] [0.1 +/- 0.01 microgram/mg protein (normal); 0.2 +/- 0.03 (transgenic)] [< 0.03 arbitrary density units (normal and transgenic)]). Serum FSH levels were increased (P < 0.05) 2 weeks after castration of normal (22 +/- 2 ng/ml) and transgenic males (135 +/- 19 ng/ml) and were suppressed (P < 0.05) by testosterone [7 +/- 0.8 ng/ml (normal); 12 +/- 2 (transgenic)] or estradiol [14 +/- 1 ng/ml (normal); 16 +/- 1 (transgenic)] replacement. The increased serum FSH levels were associated with an inverse drop (P < 0.05) in pituitary FSH content to 1 +/- 0.1 microgram/mg protein in normal and 16 +/- 2 in transgenic males. Testosterone replacement further suppressed (P < 0.05) pituitary FSH content in transgenic (3 +/- 0.5 micrograms/mg protein) but not normal (1 +/- 0.1) males.(ABSTRACT TRUNCATED AT 250 WORDS)


No overt structural or functional changes associated with PEG-coated gold nanoparticles accumulation with acute exposure in the mouse heart.
In this study, we investigated the cardiac biodistribution of polyethylene glycol (PEG)-coated AuNPs and their effects on cardiac function, structure and inflammation in both normal and cardiac remodeling mice. The model of cardiac remodeling was induced by subcutaneously injection of isoproterenol (ISO), a non-selective beta-adrenergic agonist, for 7 days. After AuNPs were injected intravenously in mice for 7 consecutive days, Au content in different organs was determined quantitatively by inductively coupled plasma mass spectrometry (ICP-MS), cardiac function and structure were measured by echocardiography, cardiac fibrosis was examined with picrosirius red staining, the morphology of cardiomyocytes was observed with hematoxylin and eosin (H & E) staining. The accumulation of AuNPs in hearts did not affect cardiac function or induce cardiac hypertrophy, cardiac fibrosis and cardiac inflammation under normal physiological condition. Cardiac AuNPs content was 6-fold higher in the cardiac remodeling mouse than normal mice. However, the increased accumulation of AuNPs in the heart did not aggravate ISO-induced cardiac hypertrophy, cardiac fibrosis or cardiac inflammation. These observations suggest that PEG-coated AuNPs possess excellent biocompatibility under both physiological and pathological conditions. Thus, AuNPs may be safe for cardiac patients and hold great promise for further development for various biomedical applications.


Sequence-recognition and cleavage of DNA by a netropsin-phenazine-di-N-oxide conjugate.
We report the synthesis, DNA-binding and cleaving properties, and cytotoxic activities of R-128, a hybrid molecule in which a bis-pyrrolecarboxamide-amidine element related to the antibiotic netropsin is covalently tethered to a phenazine-di-N-oxide chromophore. The affinity and mode of interaction of the conjugate with DNA were investigated by a combination of absorption spectroscopy, circular dichroism, and electric linear dichroism. This hybrid molecule binds to AT-rich sequences of DNA via a bimodal process involving minor groove binding of the netropsin moiety and intercalation of the phenazine moiety. The bidentate mode of binding was evidenced by linear dichroism using calf thymus DNA and poly(dA-dT).(dA-dT). In contrast, the drug fails to bind to poly(dG-dC).poly(dG-dC), because of the obstructive effect of the guanine 2-amino group exposed in the minor groove of this polynucleotide. DNase I footprinting studies indicated that the conjugate interacts preferentially with AT-rich sequences, but the cleavage of DNA in the presence of a reducing agent can occur at different sequences not restricted to the AT sites. The main cleavage sites were detected with a periodicity of about 10 base pairs corresponding to approximately one turn of the double helix. This suggests that the cleavage may be dictated by the structure of the double helix rather than the primary nucleotide sequence. The conjugate which is moderately toxic to cancer cells complements the tool box of reagents which can be utilized to produce DNA strand scission. The DNA cleaving properties of R-128 entreat further exploration into the use of phenazine-di-N-oxides as tools for investigating DNA structure.


Effects of selenomethionine supplementation on selenium status and thyroid hormone concentrations in healthy adults.
Selenium, a potential cancer prevention agent currently being tested against prostate cancer in the Selenium and Vitamin E Cancer Prevention Trial (SELECT), plays an integral role in thyroid metabolism. The effects of long-term selenium supplementation on thyroid hormone concentrations are unknown. The objective was to investigate the effects of long-term selenium supplementation on thyroid hormone concentrations. Twenty-eight healthy adults took 200 microg selenomethionine/d for 28 mo. The thyroid hormones triiodothyronine (T3), thyroxine (T4), and thyrotropin (TSH) were measured in plasma for 4 mo before supplementation and quarterly during supplementation. The assay methods were changed midstudy; the results of the 2 methods were not comparable. Therefore, one analysis was conducted based on the results of the first method, and a second analysis was based on all of the data, adjusted for the change. Serial data collection permitted a test for trends rather than simply a difference between initial and final values. By 9 mo, mean (+/-SEM) plasma selenium concentrations had increased from 1.78 +/- 0.07 micromol/L at baseline to 2.85 +/- 0.11 micromol/L for men and from 1.64 +/- 0.04 to 3.32 +/- 0.1.2 micromol/L for women. T3 concentrations in men increased 5% per year (P = 0.01). T4 and TSH concentrations were unchanged. Selenium supplementation produced no clinically significant changes in thyroid hormone concentrations. A small but statistically significant increase in T3 concentrations was noted in men, with no corresponding decreases in TSH. A subset of SELECT subjects might be monitored periodically for changes during long-term selenium supplementation.


Sexual arousal in male pigtailed monkeys (Macaca nemestrina): effects of serial matings by two males.
Male sexual arousal was examined in pigtailed monkeys (Macaca nemestrina), with the postejaculatory interval (PEI) used as the dependent variable. In male-female pairs, the PEI is shortened by 60% of normal (control) values if the male observes another male copulating with his female partner. This result demonstrates that the normal PEI is not an absolute male refractory period; males are capable of copulating sooner, and they will do so when presented with certain behavioral stimuli. A possible functional interpretation of the result with regard to male reproductive competition is presented.


Intraosseous ganglion of the lunate.
Intraosseous ganglia are occasionally found affecting the carpal bones of the hand and should be considered in the differential diagnosis of chronic wrist pain. They have characteristic radiographic findings of a cyst with a thin sclerotic rim. This case report illustrates an unusual presentation of a cyst in the lunate with successful treatment. Standard of care includes curettage and bone grafting with minimal recurrence reported.


Effect of a pelvic wedge and belt on the medial and lateral hamstring muscles during knee flexion.
[Purpose] This study developed a pelvic wedge and belt and investigated their effects on the selective activation of medial and lateral hamstring muscles during knee flexion. [Subjects and Methods] Nine adults were enrolled. The participants performed exercises without and with the pelvic wedge and belt, and the electromyographic activities of the medial and lateral hamstring muscles were recorded. [Results] The activity of the medial hamstring was increased significantly when using the pelvic wedge and belt, while the activity of the lateral hamstring did not differ significantly. [Conclusion] The pelvic wedge and belt provide a self-locked position during knee flexion in the prone position. Prone knee flexion in this position is an effective self-exercise for balanced strengthening of the medial hamstring.


Diagnosis of meniscal tears of the knee with MR imaging: effect of observer variation and sample size on sensitivity and specificity.
A wide range in the efficacy of MR imaging for the diagnosis of meniscal tears of the knee has been reported. To evaluate two possible causes for this variation, we studied how sensitivity and specificity are affected when different observers and sample sizes are used. Two hundred MR examinations of the knee in patients for whom the results of arthroscopy were available were used for the study. One hundred eight medial meniscal tears and 58 lateral meniscal tears were found at arthroscopy. The sensitivity and specificity for detection of meniscal tears were determined for the original interpretations and retrospective evaluations by three observers. Comparisons were also made between sample sizes of 25 and 100. chi 2 analysis was used for unmatched data sets and McNemar's statistic was used for matched sets. For the 200 examinations, the sensitivity was 0.89-0.93 for medial meniscal tears and 0.79-0.83 for lateral meniscal tears. The specificity was 0.86 for medial meniscal tears and 0.90-0.92 for lateral meniscal tears. Sensitivity and specificity varied widely among different observers and different sample sizes. However, we found no significant difference between any of the comparisons at the p < .05 level. The largest interobserver variation occurred in the detection of lateral meniscal tears, with a sensitivity of 0.71 for one observer and 0.88 for another observer (p = .16). The largest variation between sets of 100 examinations was a change in sensitivity for detection of lateral meniscal tears from 0.74 to 0.88 for the original interpretations (p = .10). For the sample sets of 25 cases, the variation was even larger, with the sensitivity for detection of lateral meniscal tears varying from 0.5 for one set to 1.0 for another. We conclude that chance variation related to sample size can cause large but not statistically significant variations in sensitivity and specificity in this setting. These variations are of sufficient magnitude to explain many of the differences in reported sensitivity and specificity for MR imaging in the diagnosis of meniscal tears. We found no significant difference in observer performance.


Phytohormones and their crosstalk in regulating stomatal development and patterning.
Phytohormones play important roles in regulating various aspects of plant growth and development as well as in biotic and abiotic stress responses. Stomata are openings on the surface of land plants that control gas exchange with the environment. Accumulating evidence shows that various phytohormones, including abscisic acid, jasmonic acid, brassinosteroids, auxin, cytokinin, ethylene, and gibberellic acid, play many roles in the regulation of stomatal development and patterning, and that the cotyledons/leaves and hypocotyls/stems of Arabidopsis exhibit differential responsiveness to phytohormones. In this review, we first discuss the shared regulatory mechanisms controlling stomatal development and patterning in Arabidopsis cotyledons and hypocotyls and those that are distinct. We then summarize current knowledge of how distinct hormonal signaling circuits are integrated into the core stomatal development pathways and how different phytohormones crosstalk to tailor stomatal density and spacing patterns. Knowledge obtained from Arabidopsis may pave the way for future research to elucidate the effects of phytohormones in regulating stomatal development and patterning in cereal grasses for the purpose of increasing crop adaptive responses.


Plasma Deposition and UV Light Induced Surface Grafting Polymerization of NIPAAm on Stainless Steel for Enhancing Corrosion Resistance and Its Drug Delivery Property.
When stainless steel is implanted in human bodies, the corrosion resistance and biocompatibility must be considered. In this study, first, a protective organic silicone film was coated on the surface of stainless steel by a plasma deposition technique with a precursor of hexamethyldisilazane (HMDSZ). Then, ultraviolet (UV) light-induced graft polymerization of N-isopropylacrylamide (NIPAAm) and acrylic acid (AAc) in different molar ratios were applied onto the organic silicone film in order to immobilize thermos-/pH-sensitive composite hydrogels on the surface. The thermo-/pH-sensitive composite hydrogels were tested at pH values of 4, 7.4 and 10 of a phosphate buffer saline (PBS) solution at a fixed temperature of 37 °C to observe the swelling ratio and drug delivery properties of caffeine which served as a drug delivery substance. According to the results of Fourier Transformation Infrared (FTIR) spectra and a potential polarization dynamic test, the silicone thin film formed by plasma deposition not only improved the adhesion ability between the substrate and hydrogels but also exhibited a high corrosion resistance. Furthermore, the composite hydrogels have an excellent release ratio of up to 90% of the absorbed amount after 8h at a pH of 10. In addition, the results of potential polarization dynamic tests showed that the corrosion resistance of stainless steel could be improved by the HMDSZ plasma deposition.


Replication of Tomato Yellow Leaf Curl Virus in Its Whitefly Vector, Bemisia tabaci.
Tomato yellow leaf curl virus (TYLCV) is a begomovirus transmitted exclusively by the whitefly Bemisia tabaci in a persistent, circulative manner. Replication of TYLCV in its vector remains controversial, and thus far, the virus has been considered to be nonpropagative. Following 8 h of acquisition on TYLCV-infected tomato plants or purified virions and then transfer to non-TYLCV-host cotton plants, the amounts of virus inside whitefly adults significantly increased (>2-fold) during the first few days and then continuously decreased, as measured by the amounts of genes on both virus DNA strands. Reported alterations in insect immune and defense responses upon virus retention led us to hypothesize a role for the immune response in suppressing virus replication. After virus acquisition, stress conditions were imposed on whiteflies, and the levels of three viral gene sequences were measured over time. When whiteflies were exposed to TYLCV and treatment with two different pesticides, the virus levels continuously increased. Upon exposure to heat stress, the virus levels gradually decreased, without any initial accumulation. Switching of whiteflies between pesticide, heat stress, and control treatments caused fluctuating increases and decreases in virus levels. Fluorescence in situ hybridization analysis confirmed these results and showed virus signals inside midgut epithelial cell nuclei. Combining the pesticide and heat treatments with virus acquisition had significant effects on fecundity. Altogether, our results demonstrate for the first time that a single-stranded DNA plant virus can replicate in its hemipteran vector. Plant viruses in agricultural crops are of great concern worldwide. Many of them are transmitted from infected to healthy plants by insects. Persistently transmitted viruses often have a complex association with their vectors; however, most are believed not to replicate within these vectors. Such replication is important, as it contributes to the virus's spread and can impact vector biology. Tomato yellow leaf curl virus (TYLCV) is a devastating begomovirus that infects tomatoes. It is persistently transmitted by the whitefly Bemisia tabaci but is believed not to replicate in the insect. To demonstrate that TYLCV is, in fact, propagative (i.e., it replicates in its insect host), we hypothesized that insect defenses play a role in suppressing virus replication. We thus exposed whitefly to pesticide and heat stress conditions to manipulate its physiology, and we showed that under such conditions, the virus is able to replicate and significantly influence the insect's fecundity.


Testosterone, social status and parental care in a cooperatively breeding bird.
The steroid hormone testosterone not only plays an important role in gamete production, but also influences social and aggressive behavior. Testosterone varies seasonally, peaking when competition for mates is high and declining during parental care. Surprisingly, little is known about how testosterone mediates social conflict and parental care behavior in highly social species like cooperative breeders, where group members compete for breeding opportunities and provide parental or alloparental care. We examined how testosterone differs across breeding roles in the tropical cooperatively breeding superb starling, Lamprotornis superbus. We determined whether testosterone was elevated in larger groups, and whether testosterone was negatively related to total levels of parental and alloparental care. We found that male breeders had higher testosterone than male helpers and female breeders and helpers during incubation. However, breeding males exhibited a significant decline in testosterone from incubation to chick rearing, and all individuals had similar levels during the chick rearing stage. Additionally, helpers-but not breeders-in large social groups had higher testosterone than those in small groups. Finally, testosterone was not correlated with nestling provisioning rates during chick rearing, suggesting that natural variation in the low levels of testosterone observed during periods of high parental care does not affect nestling provisioning. Together, these results offer insight into how testosterone is related to breeding roles, intra-group conflict, and parental care in a highly social species.


Adjustment sensitivity of horizontal rectus muscles in adjustable strabismus surgery.
A "marking suture" was used in 12 consecutive cases of adjustable strabismus surgery to determine the relationship between ocular alignment and the amount of horizontal muscle adjustment. The average change in ocular alignment was 2.55 prism diopters for each millimeter of muscle adjustment. Regression analysis revealed the relationship to be linear over the range of adjustments studied. The sensitivity of adjustment was the same whether the adjusted muscle was advanced or further recessed and whether the adjusted muscle was the medial rectus or the lateral rectus. No relationship was found between the distance of a recessed muscle from its normal insertion and the subsequent adjustment sensitivity of that muscle. This information, when used clinically in conjunction with a marking suture, can facilitate the adjustment phase of adjustable strabismus surgery.


Predictors of adolescent self-rated health. Analysis of the National Population Health Survey.
To examine what factors predict adolescents' concepts of their health. The study, based on the longitudinal National Population Health Survey, included 1,493 adolescents who were 12-19 at the time of interview. Sex, age, grade, family structure, income, disability, chronic health problems, social supports, social involvement, school/work involvement, smoking, alcohol bingeing, physical activities, Body Mass Index (BMI) and psychological health status variables were examined. Using ordinal multivariate regression, self-rated health was regressed on all predictors, which were entered in blocks hierarchically. The analyses revealed that adolescent perceptions of health are framed not only by their physical health status, but also by personal, socio-environmental, behavioural and psychological factors. Specifically, health problems, disability, age, female status, lower income, smoking, and higher BMI were associated with lower self-rated health. This study suggests that adolescent appraisals of their health are shaped by their overall sense of functioning, which includes both physical health and non-physical health dimensions.


Supramolecular photocatalysis by confinement--photodimerization of coumarins within cucurbit[8]urils.
Cucurbit[8]uril (as low as 10 mol%) acts as a supramolecular catalytic nanoreaction vessel and facilitates the photodimerization of coumarins in water leading to syn dimers. Saturation kinetics shows a sigmoidal dependence with a turnover number of 3.4 min(-1) and a Hill constant of 1.8 indicating a co-operative mechanism in the catalytic process.


Intracellular glutathione plays important roles in pyrrolizidine alkaloid clivorine-induced toxicity on L-02 hepatocytes.
ABSTRACT This study is aimed to evaluate the role of intracellular GSH in pyrrolizidine alkaloid clivorine-induced toxicity on human normal liver L-02 cells. Clivorine induced profound intracellular GSH depletion in a concentration-dependent manner. The GSH synthesis inhibitor L-Buthionine (S, R)-sulfoximine (BSO) significantly potentiated toxicity of clivorine, while GSH and GSH synthesis precursors N-Acetyl-L-cysteine (NAC) and S-adenosyl-L-methionine (SAM) protected cells against toxicity of clivorine. Overall, the results suggest that intracellular GSH plays an important role in the toxicity of clivorine on human hepatocytes.


[Communicating with patients: how good are our pamphlets and other written materials? An evaluation of the quality of the written information available to patients with asthma and diabetes in out-patients' clinics in Milan].
1) To identify the extent and the nature of information pamphlets available for asthmatic and diabetic patients in secondary care hospitals in Milan; 2) To investigate what patients think about the quality of pamphlets using a questionnaire developed ad hoc. Pamphlets have been identified by contacting 8 specialised ambulatory hospitals. The questionnaire was developed to achieve a quality rate in 3 dimensions of the materials (Style and readability, The condition and Treatment options). Patients were recluted in the same ambulatory hospital. 4 ambulatory hospitals and 40 patients agreed to participate in our survey. We identified 8 pamphlets (5 diabetes, 3 asthma). 6 of these have been promoted byfarmaceutical companies. The mean length was 31 pages and the issue most covered was treatment options (33% of pages dedicated). 30 pages (12% of the total) presented with publicity advertisements (6% direct and 6% indirect). The dimension with the best satisfaction rate was Condition, followed by Style and readability. In Milan, in secondary care asthmatic and diabetic patients, are often invited to consult pamphlets. They positively judged this material. During the study ambulatory physicians were poorly attentive about problems concerning patient access to information.


The functional organization of cutaneous low-threshold mechanosensory neurons.
Innocuous touch of the skin is detected by distinct populations of neurons, the low-threshold mechanoreceptors (LTMRs), which are classified as Aβ-, Aδ-, and C-LTMRs. Here, we report genetic labeling of LTMR subtypes and visualization of their relative patterns of axonal endings in hairy skin and the spinal cord. We found that each of the three major hair follicle types of trunk hairy skin (guard, awl/auchene, and zigzag hairs) is innervated by a unique and invariant combination of LTMRs; thus, each hair follicle type is a functionally distinct mechanosensory end organ. Moreover, the central projections of Aβ-, Aδ-, and C-LTMRs that innervate the same or adjacent hair follicles form narrow LTMR columns in the dorsal horn. These findings support a model of mechanosensation in which the activities of Aβ-, Aδ-, and C-LTMRs are integrated within dorsal horn LTMR columns and processed into outputs that underlie the perception of myriad touch sensations.


Mechanism of Short Term Fe Reduction by Roots : Evidence against the Role of Secreted Reductants.
The hypothesized role of secreted reducing compounds in Fe(III) reduction has been examined with Fe-deficient peanuts (Arachis hypogaea L. cv A124B). Experiments involved the exposure of roots to (a) different gas mixtures, (b) carbonyl cyanide m-chlorophenylhydrazone (CCCP), and (c) agents which impair membrane integrity.Removing roots from solution and exposing them to air or N(2) for 10 minutes did not result in any accumulation in the free space of compounds capable of increasing rates of Fe(III) reduction when roots were returned to solutions. On the contrary, exposing roots to N(2) decreased rates of Fe(III) reduction. CCCP also decreased rates of Fe(III) reduction.Acetic acid and ethylenediaminetetraacetic acid (disodium salt) (EDTA) impaired the integrity and function of the plasma membranes of roots of Fe-deficient peanuts. That is, in the presence of acetic acid or EDTA, there was an efflux of K(+) from the roots; K(+) ((86)Rb) uptake was also impaired. Acetic acid increased the efflux from the roots of compounds capable of reducing Fe(III). However, both acetic acid and EDTA caused rapid decreases in rates of Fe(III) reduction by the roots. In addition to peanuts, acetic acid also decreased rates of Fe(III) reduction by roots of Fe-deficient sunflowers (Helianthus annuus L. cv Sobrid) but not maize (Zea mays L. cv Garbo).These results suggest that, at least in the short term, the enhanced Fe(III) reduction by roots of Fe-deficient plants is not due to the secretion of reducing compounds.


Tyrosine hydroxylase deficiency in Taiwanese infants.
We analyzed the clinical manifestations, genetic mutations, treatment responses to L-dopa, and long-term neurologic outcomes in Taiwanese infants with tyrosine hydroxylase deficiency. From 1999 to May 2011, we enrolled six infants who had been diagnosed with tyrosine hydroxylase deficiency by identifying point mutations on the tyrosine hydroxylase gene. Two patients manifested fetal distress during the perinatal period. Four patients exhibited generalized tremor as their first observed neurologic sign at age 3 months. All presented brisk reflexes, hypokinesia, rigidity, distal chorea, and athetosis. We identified a novel missense mutation, I382T, and report on the first patient, to the best of our knowledge, with a homozygous R153X nonsense mutation. Five of six patients responded to L-dopa at a dose of 4.2-34.7 mg/kg/day combined with biperiden or selegiline or both. Long-term neurologic outcomes (median follow-up, 5 years and 10.5 months) revealed two patients demonstrated slightly low intelligence quotients, three demonstrated mild to moderate psychomotor retardation, and one died of respiratory failure. A higher dose of L-dopa, together with alternative therapies, may lead to improvements in motor function. However, several years of observation may be needed to reach definitive conclusions about neurologic outcomes.


Gemcitabine, Docetaxel, Capecitabine, Cisplatin, Irinotecan as First-line Treatment for Metastatic Pancreatic Cancer.
Treatment of advanced pancreatic cancer with a single therapeutic at a maximal dose has been largely ineffective at increasing survival. Combination therapies are commonly studied but often limited by toxicity. We previously showed that low-dose multiagent therapy with gemcitabine, docetaxel (taxotere), capecitabine (xeloda), and cisplatin (GTX-C) was safe, well tolerated, and effective (NCT01459614). Here, we hypothesize that adding irinotecan to GTX-C may improve survival with minimal toxicity. Patients with treatment-naïve metastatic pancreatic adenocarcinoma were treated with gemcitabine, docetaxel (taxotere), capecitabine (xeloda), cisplatin, and irinotecan (GTX-CI). Treatment consisted of capecitabine 500 mg twice daily on days 1-14 and gemcitabine 300 to 500 mg/m2, docetaxel 20 mg/m2, cisplatin 15 to 20 mg/m2, and irinotecan 20 to 60 mg/m2 on days 4 and 11 of a 21-day cycle. The primary objective was 9-month overall survival (OS). Secondary objectives included response rate (RR), disease control rate (DCR), progression-free survival (PFS), and OS. The regimen was well tolerated. The recommended phase II dose was gemcitabine 500 mg/m2, docetaxel 20 mg/m2, capecitabine 500 mg po bid, cisplatin 20 mg/m2, and irinotecan 20 mg/m2. Median follow-up in phase II was 11.02 months (2.37-45.17). Nine-month OS rate was 57% [95% confidence interval (CI): (41-77)]. RR was 57% [95% CI: (37-75) 50% PR and 7% CR]. DCR was 87% [95% CI: (69-96)]. Median OS and PFS were 11.02 [95% CI: (8.54-21.09)] and 8.34 [95% CI: (6.34-NA)] months, respectively. The addition of irinotecan to GTX-C was safe and well tolerated. While the study did not meet its primary objective, the responses were clinically meaningful using a well-tolerated regimen. We aimed to optimize the previously reported efficacious regimen of low-dose multiagent therapy with GTX-C for the treatment of metastatic pancreatic ductal adenocarcinoma by adding irinotecan. The primary objective was not met, but GTX-CI was well tolerated. The RR of 57%, median PFS of 8.3 months, median OS of 11 months, and 36-month OS rate of 19% suggest clinical benefit. Further optimization of this regimen is warranted.


[Idiopathic thrombocytopenic purpura].
Several recent studies have suggested the eradication of Helicobacter pylori (H. pylori) can increase the platelet count in patients with chronic idiopathic thrombocytopenic purpura (ITP). A systematic review of the literature published in English shows that overall platelet responses were observed in over 50% of the patients after successful eradication. The response rate tended to be higher in countries with a high prevalence ratio of H. pylori infection in background population, in patients with less severe degree of thrombocytopenia and in those with shorter disease duration. The detection and eradication of H. pylori should be considered as the first line of treatment in all Japanese patients with chronic ITP.


SepT, a novel protein specific to multicellular cyanobacteria, influences peptidoglycan growth and septal nanopore formation in Anabaena sp. PCC 7120.
Anabaena sp. PCC 7120 grows by forming filaments of communicating cells and is considered a paradigm of bacterial multicellularity. Molecular exchanges between contiguous cells in the filament take place through multiprotein channels that traverse the septal peptidoglycan through nanopores connecting their cytoplasms. Besides, the septal-junction complexes contribute to strengthen the filament. In search for proteins with coiled-coil domains that could provide for cytoskeletal functions in Anabaena, we identified SepT (All2460). SepT is characteristic of the phylogenetic clade of filamentous cyanobacteria with the ability to undergo cell differentiation. SepT-GFP fusions indicate that the protein is located at the cell periphery and, conspicuously, in the intercellular septa. During cell division, the protein is found at midcell resembling the position of the divisome. The bacterial adenylate cyclase two-hybrid analysis shows SepT interactions with itself and putative elongasome (MreB, RodA), divisome (FtsW, SepF, ZipN), and septal-junction (SepJ)-related proteins. Thus, SepT appears to rely on the divisome for localization at mature intercellular septa to form part of intercellular protein complexes. Two independently obtained mutants lacking SepT showed alterations in cell size and impaired septal and peripheral peptidoglycan incorporation during cell growth and division. Notably, both mutants showed conspicuous alterations in the array of nanopores present in the intercellular peptidoglycan disks, including aberrant nanopore morphology, number, and distribution. SepT appears, therefore, to be involved in the control of peptidoglycan growth and the formation of cell-cell communication structures that are at the basis of the multicellular character of this group of cyanobacteria. IMPORTANCE Multicellular organization is a requirement for the development of complex organisms, and filamentous cyanobacteria such as Anabaena represent a paradigmatic case of bacterial multicellularity. The Anabaena filament can include hundreds of communicated cells that exchange nutrients and regulators and, depending on environmental conditions, can include different cell types specialized in distinct biological functions. Hence, the specific features of the Anabaena filament and how they are propagated during cell division represent outstanding biological issues. Here, we studied SepT, a novel coiled-coil-rich protein of Anabaena that is located in the intercellular septa and influences the formation of the septal specialized structures that allow communication between neighboring cells along the filament, a fundamental trait for the performance of Anabaena as a multicellular organism.


Predicting membrane protein types with bragging learner.
The membrane protein type is an important feature in characterizing the overall topological folding type of a protein or its domains therein. Many investigators have put their efforts to the prediction of membrane protein type. Here, we propose a new approach, the bootstrap aggregating method or bragging learner, to address this problem based on the protein amino acid composition. As a demonstration, the benchmark dataset constructed by K.C. Chou and D.W. Elrod was used to test the new method. The overall success rate thus obtained by jackknife cross-validation was over 84%, indicating that the bragging learner as presented in this paper holds a quite high potential in predicting the attributes of proteins, or at least can play a complementary role to many existing algorithms in this area. It is anticipated that the prediction quality can be further enhanced if the pseudo amino acid composition can be effectively incorporated into the current predictor. An online membrane protein type prediction web server developed in our lab is available at http://chemdata.shu.edu.cn/protein/protein.jsp.


Transition State Models for Understanding the Origin of Chiral Induction in Asymmetric Catalysis.
In asymmetric catalysis, a chiral catalyst bearing chiral center(s) is employed to impart chirality to developing stereogenic center(s). A rich and diverse set of chiral catalysts is now available in the repertoire of synthetic organic chemistry. The most recent trends point to the emergence of axially chiral catalysts based on binaphthyl motifs, in particular, BINOL-derived phosphoric acids and phosphoramidites. More fascinating ideas took shape in the form of cooperative multicatalysis wherein organo- and transition-metal catalysts are made to work in concert. At the heart of all such manifestations of asymmetric catalysis, classical or contemporary, is the stereodetermining transition state, which holds a perennial control over the stereochemical outcome of the catalytic process. Delving one step deeper, one would find that the origin of the stereoselectivity is delicately dependent on the relative stabilization of one transition state, responsible for the formation of the predominant stereoisomer, over the other transition state for the minor stereoisomer. The most frequently used working hypothesis to rationalize the experimentally observed stereoselectivity places an undue emphasis on steric factors and tends to regard the same as the origin of facial discrimination between the prochiral faces of the reacting partners. In light of the increasing number of asymmetric catalysts that rely on hydrogen bonding as well as other weak non-covalent interactions, it is important to take cognizance of the involvement of such interactions in the sterocontrolling transition states. Modern density functional theories offer a pragmatic and effective way to capture non-covalent interactions in transition states. Aided by the availability of such improved computational tools, it is quite timely that the molecular origin of stereoselectivity is subjected to more intelligible analysis. In this Account, we describe interesting molecular insights into the stereocontrolling transition states of five reaction types, three of which provide access to chiral quaternary carbon atoms. While each reaction has its own utility and interest, the focus of our research has been on the mechanism and the origin of the enantio- and diastereoselectivity. In all of the examples, such as asymmetric diamination, sulfoxidation, allylation, and Wacker-type ring expansion, the role played by non-covalent interactions in the stereocontrolling transition states has been identified as crucial. The transfer of the chiral information from the chiral catalyst to the product is identified as taking place through a series of non-covalent interactions between the catalyst and a given position/orientation of the substrate in the chiral environment offered by the axially chiral catalyst. The molecular insights enunciated herein allude to abundant opportunities for rational modifications of the present generation of catalysts and the choice of substrates in these as well as related families of reactions. It is our intent to propose that the domain of asymmetric catalysis could enjoy additional benefits by having knowledge of the vital stereoelectronic interactions in the stereocontrolling transition states.


Education-based approach to addressing non-evidence-based practice in preventing NSAID-associated gastrointestinal complications.
To evaluate an evidence-based educational program for improving strategies for prevention of nonsteroidal anti-inflammatory drug (NSAID)-associated gastrointestinal (GI) complications. Four hundred and fifty-six specialists replied to a questionnaire that covered issues related to NSAID-induced adverse effects. They also collected data from their last five consecutive patients before and after they had attended an evidence-based seminar on GI prevention strategies. Four hundred and forty-one of 456 specialists (96.7%) participated in the survey, and 382 (83.7%) in the education-based study that recorded data from 3728 patients. The specialists overestimated the risk of GI complications with NSAIDs, underestimated the GI safety profile of coxibs, but were aware of the risk factors and of the current prevention strategies. Proton pump inhibitors were co-prescribed with NSAIDs in > 80% of patients with and without risk factors. The educational program had little impact on prescribing habits. Specialists are informed of advances in NSAID-associated adverse effects and have high rates of GI-prevention therapy. Our educational program did not alter these rates.


Regulation of arabinose and xylose metabolism in Escherichia coli.
Bacteria such as Escherichia coli will often consume one sugar at a time when fed multiple sugars, in a process known as carbon catabolite repression. The classic example involves glucose and lactose, where E. coli will first consume glucose, and only when it has consumed all of the glucose will it begin to consume lactose. In addition to that of lactose, glucose also represses the consumption of many other sugars, including arabinose and xylose. In this work, we characterized a second hierarchy in E. coli, that between arabinose and xylose. We show that, when grown in a mixture of the two pentoses, E. coli will consume arabinose before it consumes xylose. Consistent with a mechanism involving catabolite repression, the expression of the xylose metabolic genes is repressed in the presence of arabinose. We found that this repression is AraC dependent and involves a mechanism where arabinose-bound AraC binds to the xylose promoters and represses gene expression. Collectively, these results demonstrate that sugar utilization in E. coli involves multiple layers of regulation, where cells will consume first glucose, then arabinose, and finally xylose. These results may be pertinent in the metabolic engineering of E. coli strains capable of producing chemical and biofuels from mixtures of hexose and pentose sugars derived from plant biomass.


The novel cytochrome c6 of chloroplasts: a case of evolutionary bricolage?
Cytochrome c6 has long been known as a redox carrier of the thylakoid lumen of cyanobacteria and some eukaryotic algae that can substitute for plastocyanin in electron transfer. Until recently, it was widely accepted that land plants lack a cytochrome c6. However, a homologue of the protein has now been identified in several plant species together with an additional isoform in the green alga Chlamydomonas reinhardtii. This form of the protein, designated cytochrome c6A, differs from the 'conventional' cytochrome c6 in possessing a conserved insertion of 12 amino acids that includes two absolutely conserved cysteine residues. There are conflicting reports of whether cytochrome c6A can substitute for plastocyanin in photosynthetic electron transfer. The evidence for and against this is reviewed and the likely evolutionary history of cytochrome c6A is discussed. It is suggested that it has been converted from a primary role in electron transfer to one in regulation within the chloroplast, and is an example of evolutionary 'bricolage'.


Post-traumatic stress disorder in bam-survived students who immigrated to Kerman, four months after the earthquake.
The 6.3 Richter earthquake in Bam killed 26,000 and injured 30,000 people, and destroyed 85% of houses according to the literature. Children are affected more than others by disasters. In this study, we assessed the rate of post-traumatic stress disorder and psychological problems in survived Bam students in Kerman four months after the earthquake and their relations with demographic factors, family loss, house destruction, and body injury. A group of 433 students attended this cross sectional study. Watson post traumatic stress disorder questionnaire and Symptom Checklist-90 were applied for students older than 15 years of age and Yule post-traumatic stress disorder and Rutter (parents' version) questionnaires were used for students younger than 15. Multiple regression analysis was used to assess the effects of variables on psychopathologies. Post-traumatic stress disorder was detected in 36.3% of the students older than 15 years of age and 51.6% of the students younger than 15. Behavioral problems were present in 31.3% of the children and the mean score of Global Symptom Index was 1.13+/-0.59. Body injury and living with family were correlated with post-traumatic stress disorder and Symptom Checklist-90 items in students older than 15. Female sex was correlated with post-traumatic stress disorder and behavioral problems in students younger than 15. The prevalence of post-disaster post-traumatic stress disorder was high in children. Younger age, female gender, living apart from family, geographic living conditions, being in the center of stress or not after the earthquake, body injury, and loss of family members form the basis for developing post-disaster psychopathology. Due to incomplete presence of post-traumatic stress disorder criteria or high rate of overlooking post-traumatic stress disorder in children, the disorder may be misdiagnosed or remain undiagnosed in them, which warrants more consideration.


Attrition Rates and Incidence of Mental Health Disorders in an Attention-Deficit/Hyperactivity Disorder (ADHD) Cohort, Active Component, U.S. Armed Forces, 2014-2018.
Attention-deficit/hyperactivity disorder (ADHD) is a common childhood diagnosis and affects the pool of potential military applicants. Early detection and treatment of ADHD may decrease the risk of developing comorbidities; however, accession policy in place during this study period (2014-2018) disqualified applicants who used ADHD medication for more than 24 months cumulative after age 14. The objective of this study was to assess attrition from military service in newly accessed active component service members diagnosed with ADHD as compared to controls. In addition, attrition rates and incidence rates of mental health diagnoses were assessed in service members with ADHD by treatment status (i.e., treated vs untreated ADHD) where treatment was defined as being dispensed an FDA-approved ADHD medication at least twice within 181 days. Almost two-thirds (64.8%) of newly accessed ADHD cases in 2014 were identified after enlistment medical screening at Military Entrance Processing Stations (MEPS) (i.e., post-MEPS). These post-MEPS ADHD cases accounted for 99.1% of the treated ADHD cases. The vast majority of treated cases (91.0%) were dispensed ADHD medication within 6 months of accession. The treated ADHD group had higher rates of attrition and incidence of mental health disorders during the followup period. These study findings highlight the problem of nondisclosure of ADHD among military applicants. Future changes to enlistment standards should consider the optimal way to promote applicant disclosure of ADHD during MEPS screening or for medical waiver review and should discourage withholding an ADHD diagnosis during enlistment.


Canes may not improve spatiotemporal parameters of walking after stroke: a systematic review of cross-sectional within-group experimental studies.
To examine whether using a cane would improve spatiotemporal parameters of walking, i.e., speed, stride length, cadence, and symmetry after stroke. Searches were conducted in eight databases. The experimental condition was walking with a cane. Four outcomes were of interest: walking speed, stride length, cadence, and symmetry. Twelve studies were included. Results from nine studies suggested that individuals with stroke walked 0.01 m/s (SD 0.06) slower with a single-point cane, compared with no cane. Two studies suggested a reduction in cadence (MD-5 steps/min, SD2) and an increase in stride length (MD 0.08 m, SD 0.01). Three studies suggested that individuals walked 0.06 m/s (SD 0.07) slower with a four-point cane, compared with no cane. Four studies suggested that individuals walked 0.06 m/s (SD 0.04) faster with a single- point cane compared with a four-point cane. Results regarding other outcomes were inconclusive. Results showed no worthwhile improvements in spatiotemporal parameters of walking with a single-point cane and a slight reduction with a four-point cane, compared with no cane. Individuals walked slightly faster with a single-point cane compared with a four-point cane, but the evidence is insufficient to support this superiority.IMPLICATIONS FOR REHABILITATIONA single-point cane may not improve spatiotemporal parameters of walking after stroke.Walking with a four-point cane may slightly decrease spatiotemporal parameters of walking.Canes may be prescribed without the fear of negatively impairing walking kinematics.


Regulation of hepatic glutamate metabolism. Role of 2-oxoacids in glutamate release from isolated perfused rat liver.
In isolated perfused rat liver, addition of the oxoanalogues of leucine, isoleucine, methionine and phenylalanine is followed by a rapid and reversible stimulation of glutamate release. This is not observed with the corresponding amino acids or 2-oxoisovalerate, 2-oxoglutarate or oxaloacetate. The increased glutamate release by the liver is accompanied by a decrease in the tissue contents of 2-oxoglutarate and glutamate by about 25% and 50%, respectively. During the metabolism of glutamine, i.e. conditions with elevated tissue glutamate concentrations, 2-oxoacid-induced glutamate release is stimulated. In the presence of glutamine (5 mM), 2-oxoisocaproate, 2-oxo-4-methylvalerate and 2-oxo-4-methylthiobutyrate were found to be most effective and glutamate release by the liver increased linearly from about 80 nmol g-1 min-1 to 600 nmol g-1 min-1 at increasing 2-oxoacid concentrations up to 1 mM. When glutamate tissue levels were decreased by phenylephrine, stimulation of glutamate release by 2-oxoisocaproate was markedly diminished. 2-Oxoacid-stimulated glutamate release is independent of oxoacid metabolism, indicating that the effect is probably not explained by a 2-oxoacid/glutamate exchange across the liver plasma membrane. 2-Oxoacid-induced glutamate export predominantly occurs in a sodium-independent way. At low concentrations of 2-oxoisocaproate (below 0.2 mM), the increased glutamate release was accompanied by a slight inhibition of 14CO2 production from added [14C]glutamate, indicating a simultaneous glutamate uptake and release also under these conditions. Stimulation of glutamate release by 2-oxoisocaproate is followed by a decreased rate of urea and glutamine synthesis from portal ammonia, as a consequence of an increased glutamate release.


A peer-led mobile outreach program and increased utilization of detoxification and residential drug treatment among female sex workers who use drugs in a Canadian setting.
The objectives of this study were to examine the determinants of using a peer-led mobile outreach program (the Mobile Access Project [MAP]) among a sample of street-based female sex workers (FSWs) who use drugs in an urban Canadian setting and evaluate the relationship between program exposure and utilizing addiction treatment services. A detailed questionnaire was administered at baseline and bi-annual follow-up visits over 18 months (2006-2008) to 242 FSWs in Vancouver, Canada. We used bivariate and multivariate logistic regression with generalized estimating equations for both objectives, reporting unadjusted and adjusted odds ratios (AOR) with 95% confidence intervals (CIs). Over 18 months, 42.2% (202) reports of peer-led mobile outreach program use were made. High-risk women, including those servicing a higher weekly client volume (10+ compared to <10; AOR: 1.7, 95%CIs: 1.1-2.6) and those soliciting clients in deserted, isolated settings (AOR: 1.7, 95%CIs: 1.1-2.7) were more likely to use the program. In total, 9.4% (45) reports of using inpatient addiction treatment services were made (7.5% detoxification; 4.0% residential drug treatment), and 33.6% (161) using outpatient treatment (28.8% methadone; 9.6% alcohol/drug counsellor). Women who used the peer-led mobile outreach were more likely to use inpatient addiction treatment (AOR: 4.2, 95%CIs: 2.1-8.1), even after adjusting for drug use, environmental-structural factors, and outpatient drug treatment. Our findings demonstrate that FSWs at higher risk for sexually transmitted infections and violence are more likely to access this peer-led mobile outreach program and suggest that the program plays a critical role in facilitating utilization of detoxification and residential drug treatment.


Febrile-range hyperthermia accelerates caspase-dependent apoptosis in human neutrophils.
Human neutrophilic polymorphonuclear leukocytes (PMNs) are central to innate immunity and are responsible for clearance of pathogens. PMNs undergo a tightly regulated apoptosis program that allows for timely clearance of PMNs without extravasation of toxic intracellular contents. We investigated the rate of spontaneous apoptosis of human peripheral blood PMNs cultured at basal (37 degrees C) and febrile-range (39.5 degrees C) temperatures (FRT). We found that PMN apoptosis is accelerated at FRT, reaching approximately 90% completion by 8 h at 39.5 degrees C vs 18 h at 37 degrees C based on morphologic criteria. Caspase-8 activation peaked within 15 min of PMN exposure to FRT, and subsequent activation of caspase-3 and -9, cleavage of the BH3 (Bcl-2 homology domain 3) only protein Bid, and mitochondrial release of cytochrome c were also greater in FRT-exposed PMNs. Inhibition of caspase-3, -8, and -9 conferred comparable protection from apoptosis in FRT-exposed PMNs. These results demonstrate that exposure to FRT enhances caspase-8 activation and subsequent mitochondrial-dependent and mitochondrial-independent apoptosis pathways. The PMN survival factors G-CSF, GM-CSF, and IL-8 each prolonged PMN survival at 37 degrees C and 39.5 degrees C, but did not reduce the difference in survival at the two temperatures. In a mouse model of intratracheal endotoxin-induced alveolitis, coexposure to FRT (core temperature approximately 39.5 degrees C) doubled the proportion of bronchoalveolar PMNs undergoing apoptosis compared with euthermic mice. This process may play an important role in limiting inflammation and tissue injury during febrile illnesses.


Potential distribution of an invasive pest, Euplatypus parallelus, in China as predicted by Maxent.
Euplatypus parallelus is a highly polyphagous invasive pest native to Central and South America. In recent years it has invaded many countries in Africa and Asia and resulted in considerable economic loss. In China it has been reported to have invaded Taiwan, and been also recorded in Hainan Province. Until now there has been no invasion into the mainland. In order to better manage this invasive pest, here we predicted the suitable area of E. parallelus in China by the Maxent model. The Maxent model predicted the potential distribution of E. parallelus with a test AUC value of 0.962 and a training AUC value of 0.978. Temperature seasonality (Bio_04), annual temperature range (Bio_07), annual precipitation (Bio_12) and mean temperature of the coldest quarter (Bio_11) were the strongest predictors of E. parallelus distribution with 32.1%, 19.8%, 15% and 10.4% contributions, respectively. The potential suitable area for E. parallelus was mainly distributed in the southeastern coast, the southwestern border, and Taiwan and Hainan provinces in China, and the highly suitable areas were located in the northern coast of Hainan Province and the southwestern coast of Taiwan Province. This pest prefers a stable, warm and rainy climate, which indicates that tropics and subtropics would be its ideal area. Euplatypus parallelus has invaded Hainan and Taiwan in China. Measures should be taken to prevent it from spreading on these two islands. Moreover, strict quarantine, biological study and control measures are necessary to block its spread, invasion and damage, especially in these climate-suitable areas. © 2018 Society of Chemical Industry.


Fizeau-type interferometric probe to measure geometrical thickness of silicon wafers.
We developed an optical interferometric probe for measuring the geometrical thickness and refractive index of silicon wafers based on a Fizeau-type spectral-domain interferometer, as realized by adopting the optical fiber components of a circulator and a sheet-type beam splitter. The proposed method enables us to achieve a much simpler optical composition and higher immunity to air fluctuations owing to the use of fiber components and a common-path configuration as compared to a bulk-type optical configuration. A femtosecond pulse laser having a spectral bandwidth of 80 nm at a center wavelength of 1.55 µm and an optical spectrum analyzer having a wavelength uncertainty of 0.02 nm were used to acquire multiple interference signals in the frequency domain without a mechanical phase-shifting process. Among the many peaks in the Fourier-transformed signals of the measured interferograms, only three interference signals representing three different optical path differences were selected to extract both the geometrical thickness and group refractive index of a silicon wafer simultaneously. A single point on a double-sided polished silicon wafer was measured 90 times repetitively every two seconds. The geometrical thickness and group refractive index were found to be 476.89 µm and 3.6084, respectively. The measured thickness is in good agreement with that of a contact type method within the expanded uncertainty of contact-type instruments. Through an uncertainty evaluation of the proposed method, the expanded uncertainty of the geometrical thickness was estimated to be 0.12 µm (k = 2).


Defining the current role of immune checkpoint inhibitors in the treatment of mismatch repair-deficient/microsatellite stability-high colorectal cancer and shedding light on future approaches.
Introduction: Mismatch repair deficient (MMR-D)/microsatellite instability-high (MSI-H) colorectal cancer (CRC) carries unique biologic features including high tumor mutation burden, increased amount of mutation-associated neoantigen generation, and the presence of marked tumor-infiltrating lymphocytes. Immune checkpoint inhibitor (ICI) therapy has rapidly changed the treatment algorithm of MMR-D/MSI-H CRC.Areas covered: In this review article, we discuss the recent data regarding the use of ICIs in metastatic MMR-D/MSI-H CRC patients. We also elaborated on potential biomarkers of ICI response and innovative therapeutic approaches that may prevail resistance mechanisms for the treatment of MMR-D/MSI-H colorectal cancer.Expert opinion: Pembrolizumab was recently granted approval by the FDA as first-line therapy for metastatic MMR-D/MSI-H CRC based on the results of the Keynote 177 study. The combination of nivolumab and ipilimumab will also likely be a choice for the initial therapy of MMR-D/MSI-H CRC in the near future. More therapeutic modalities with novel immunomodulatory agents as well as targeted therapy directed to immune resistance pathways are needed. The novel approaches discussed in this review article will define potential treatment options for the management of MMR-D/MSI-H CRC patients who progress on first-line ICI therapy.


[Effects of Canalization on the Iron Deposition in Sanjiang Plain].
Canalization is the representative process and landscape of wetland reclamation. A typical ditch system of four levels near the Honghe National Nature Reserve in Sanjiang Plain was selected. Deposition plates were set on the sediments along the ditch level and the remained natural wetland nearby was quantitatively sampled for two years as the control. The deposition fluxes, total iron concentration, iron oxides and their components, as well as biogenic elements in the sediments collected by deposition plates were measured. The results showed that the litter, mud/sand and total deposition fluxes showed no significant differences between different ditch levels, with the means of (57.00 ± 16.90) g x (m2 x a)(-1), (3 997.57 ± 798.98) g x (m2 x a)(-1) and (4054.57 ± 792.91) g x (m2 x a)(-1), respectively. The litter flux decreased with the increase of ditch level, and the flux in the natural wetland [ (120.26 ± 19.42) g x (m2 x a)(-1) ] was significantly greater than that of the ditches. The mud/sand [ (35.41 ± 11.15 ) g x (m2 x a)(-1)] and total deposition fluxes [ (155.67 ± 20.75) g x ( m2 x a](-1) ] were significantly smaller than those of the ditches. There were no significant differences in the total iron between different ditches and natural wetland, while the free iron oxide content in the ditch sediments was significantly lower than that of natural wetland sediment. Except for the main ditch, the amorphous and complex iron oxides in the other ditch and natural wetland sediments showed no significant differences. The free degree of the iron oxide in ditch sediments was 60.2% of that in the natural wetland, while the differences in the complex degree and the activated degree were insignificant. The differences in the total organic carbon, total nitrogen and total phosphorus were insignificant, and all were smaller than those of the natural wetland, with the percentages of 14.6%, 31.6% and 41.0%, respectively. It could be concluded that the effects of canalization on iron and biogenic elements were significant. Consequently, rational agricultural water managements are strongly recommended to avoid the potential environmental and ecological risks caused by canalization in Sanjiang Plain.


Dye-Sensitized Hydrobromic Acid Splitting for Hydrogen Solar Fuel Production.
Hydrobromic acid (HBr) has significant potential as an inexpensive feedstock for hydrogen gas (H2) solar fuel production through HBr splitting. Mesoporous thin films of anatase TiO2 or SnO2/TiO2 core-shell nanoparticles were sensitized to visible light with a new RuII polypyridyl complex that served as a photocatalyst for bromide oxidation. These thin films were tested as photoelectrodes in dye-sensitized photoelectrosynthesis cells. In 1 N HBr (aq), the photocatalyst undergoes excited-state electron injection and light-driven Br- oxidation. The injected electrons induce proton reduction at a Pt electrode. Under 100 mW cm-2 white-light illumination, sustained photocurrents of 1.5 mA cm-2 were measured under an applied bias. Faradaic efficiencies of 71 ± 5% for Br- oxidation and 94 ± 2% for H2 production were measured. A 12 μmol h-1 sustained rate of H2 production was maintained during illumination. The results demonstrate a molecular approach to HBr splitting with a visible light absorbing complex capable of aqueous Br- oxidation and excited-state electron injection.


[Specific and nonspecific (anti-IgE) immunotherapy of allergic diseases].
Specific immunotherapy comprises a special form of allergy treatment which consists of stepwise increasing doses of the allergen, given subcutaneously or orally with the aim to reprogram the specific immunity (from allergy to tolerance). This requires some experience and an exact allergological workup, since the main mistake in the treatment of this specific immunotherapy is the selection of unsuitable patients. The effectivity of specific immunotherapy is well documented for bee and wasp venom allergy, pollinosis and more and more also for asthma bronchiale. A type of unspecific passive immunotherapy is the injection of anti-IgE antibodies. However they block only soluble IgE and are not able to remove mast cell bound IgE. Thus, the allergic reactivity persists. Clinical studies have shown some efficacy in asthma as well as in pollinosis. However, at present it is unclear, whether this drug, which has to be given every two to four weeks in relatively high concentrations, is effective in patients with polysensitization (i.e. patients suffering from mite, fungal and pollen allergy) or is effective in patients which have manifestations of their allergy in different organs.


Broad Dissemination of Plasmids across Groundwater-Fed Rapid Sand Filter Microbiomes.
Biological rapid sand filtration is a commonly employed method for the removal of organic and inorganic impurities in water which relies on the degradative properties of microorganisms for the removal of diverse contaminants, but their bioremediation capabilities vary greatly across waterworks. Bioaugmentation efforts with degradation-proficient bacteria have proven difficult due to the inability of the exogenous microbes to stably colonize the sand filters. Plasmids are extrachromosomal DNA elements that can often transfer between bacteria and facilitate the flow of genetic information across microbiomes, yet their ability to spread within rapid sand filters has remained unknown. Here, we examine the permissiveness of rapid sand filter communities toward four environmentally transmissible plasmids, RP4, RSF1010, pKJK5, and TOL (pWWO), using a dual-fluorescence bioreporter platform combined with fluorescence-activated cell sorting (FACS) and 16S rRNA gene amplicon sequencing. Our results reveal that plasmids can transfer at high frequencies and across distantly related taxa from rapid sand filter communities, emphasizing their potential suitability for introducing bioremediation determinants in the microbiomes of underperforming water purification plants. IMPORTANCE The supply of clean water for human consumption is being challenged by the appearance of anthropogenic pollutants in groundwater ecosystems. Because many plasmids can transfer horizontally between members of bacterial communities, they comprise promising vectors for the dissemination of pollutant-degrading genetic determinants within water purification plants. However, their ability to spread within groundwater-fed rapid sand filters has not been explored. Here, we investigate the transfer dynamics of four transmissible plasmids across rapid sand filter communities originating from three different waterworks in Denmark. Our results revealed a significant ability of natural plasmids to transfer at high frequencies and across distantly related taxa in the absence of plasmid selection, indicating their potential suitability as vectors for the spread of bioremediation determinants in water purification plants. Future work is required to assess the biotechnological applicability and long-term maintenance of exogenous plasmids within sand filter communities.


Liver resection with normothermic ischaemia exceeding 1 h.
A retrospective study was made of 34 patients who underwent major liver resection with a single period of vascular occlusion exceeding 60 min. The liver remnant was normal in all cases. Vascular occlusion was achieved by continuous portal triad clamping (15 patients), hepatic vascular exclusion (15) or a sequential combination of both procedures (four). Liver cooling was not used. The mean (s.e.m.) duration of continuous normothermic liver ischaemia was 73.6 (2.5) (range 60-127) min. The mean (s.e.m.) amount of blood transfused during operation was 5.3(0.8) units packed red cells. There were no deaths after surgery and the postoperative course was uneventful, or limited to asymptomatic pleural effusion, in 18 patients. Three patients suffered postoperative bleeding necessitating further surgery and one of these required reintervention for a prolonged bile leak. Four patients had transient liver failure that resolved spontaneously within 15 days. There was a 13-fold increase in serum transaminase activities and the proaccelerin level was 45 per cent that of normal on day 1 after operation. These changes were returning to normal levels within 15 days. Continuous vascular occlusion during major liver resection is a useful manoeuvre that may be performed safely on normal hepatic parenchyma for up to 90 min.


Dual-Layer All-Textile Flexible Pressure Sensor Coupled by Silver Nanowires with Ti3C2-Mxene for Monitoring Athletic Motion during Sports and Transmitting Information.
At present, wearable flexible pressure sensors have broad application prospects in fields such as motion monitoring and information transmission. However, it is still a challenge to design flexible pressure sensors with high sensitivity over a large sensing range and simple fabrication. Here, we use a simple "dipping-drying" method to fabricate a fabric-based flexible pressure sensor by coupling silver nanowires (AgNWs) with Ti3C2-MXene. The interaction between MXene and AgNWs helps realize a dual-layer sensing network, achieving good synergistic effects between pressure sensitivity and sensing range. The effects of the material combination and dip-coating sequence on the sensor's performance are systematically studied. The results show that the sensor was impregnated sequentially with AgNWs solution, and the MXene solution has the highest sensitivity (0.168 kPa-1) over a wide range (190 kPa). Meanwhile, it has the advantages of low response hysteresis and detection limit, as well as good linearity and durability. We further demonstrate the application of this sensor in human physiological signal monitoring and motion pattern recognition. It can also encrypt and transmit information according to different pressing states. In addition, the proposed pressure sensor array exhibits spatial resolution detection capabilities, laying the foundation for applications in the fields of motion monitoring and human-computer interaction.


Effect of Ipragliflozin (ASP1941), a novel selective sodium-dependent glucose co-transporter 2 inhibitor, on urinary glucose excretion in healthy subjects.
Hyperglycaemia is associated with serious complications, significant morbidity and death. Despite the availability of a wide range of therapeutic options, many patients with diabetes mellitus fail to achieve or maintain recommended glycaemic goals. Ipragliflozin (ASP1941) is a novel, selective inhibitor of the sodium-dependent glucose co-transporter 2, which is highly expressed in the proximal tubules of the kidneys. It suppresses renal glucose reabsorption and increases urinary glucose excretion (UGE), potentially providing an insulin-independent treatment option for type 2 diabetes. This multiple ascending-dose study assessed the safety, tolerability, pharmacokinetics and pharmacodynamics of ipragliflozin in healthy subjects after single doses and multiple once-daily doses for 10 days (dose levels: 5-600 mg). Ipragliflozin was well tolerated following single and multiple once-daily oral dosing. Ipragliflozin was rapidly absorbed with a median time to reach the maximum plasma concentration of 1.3 hours after the last dose. The area under the plasma concentration-time curve increased proportionally with increasing dose. The mean elimination half-life was 12 hours following the last dose. Ipragliflozin dose dependently increased UGE up to a maximum of approximately 59 g (327 mmol) of glucose excreted over 24 hours following multiple doses, without affecting plasma glucose levels in healthy subjects. Administration of ipragliflozin was well tolerated and resulted in a rapid, dose-dependent increase in glucosuria. Pharmacodynamic and pharmacokinetic data suggest that ipragliflozin is suitable for prolonged once-daily oral treatment.


The effect of a milk diet on the retention of 74As labelled arsenic in mice.
The effect of 3 diets with different proportion of milk proteins on retention of arsenic was studied in mice. Arsenic was administered via drinking water in concentration 50 mg As [III] per litre labelled with 74As in amount 2.96 MBq per 100 ml. After 2, 4, 8, 16 and 32 days of exposure the mice in groups of 6 each from all 3 experimental cohorts were decapitated and the content of 74As was determined in whole body, blood, liver, kidneys, spleen, lungs and heart by measuring of the activity in Gamma Scintillation Counter Tesla. From the intake of drinking water and amount of arsenic found in the experimental animals was calculated retention of arsenic in mice at different exposure intervals in all three diets. The values of arsenic found at the exposure intervals in examined materials of all three experimental cohorts were compared. The results obtained indicate that a milk diet has no adverse effect at exposure to arsenic in the sense of enhancing of its retention in mice at given experimental conditions. The found data seemed to suggest that the milk protein rich diet caused retardation of the increase of arsenic concentrations in blood, liver and kidneys that might lead at a lower exposure rate to a decrease in arsenic content in tissues of exposed animals.


Formulation and evaluation of insulin dry powder for inhalation.
Dry powder formulation of insulin for pulmonary administration was prepared to obtain increased drug deposition in the alveolar absorptive region. The deposition was studied by investigating the dispersion and deaggregation of insulin from the carrier lactose using an Andersen cascade impactor and twin stage impinger. The subsequent absorption following the deposition was studied by in vivo method. Insulin in solution with absorption promoters was lyophilized. The powder was incorporated with lactose of different grades and their combinations as carriers to deliver using an inhaler device. Solid-state characteristics of the carrier as well as the drug powder were assessed by particle size and distribution measurement. The flow properties such as moisture content, powder density, angle of repose, and carr's compressibility index of the powder mixture were determined. The aerosol behavior of the powder was studied by dispersion using rotahaler(c) connected to a twin-stage impinger (TSI) and an eight-stage Andersen cascade impactor (ACI) operating at different flow rates of 30-90 l/min. The in vivo performance was studied by deliverance to the respiratory tract of guinea pigs. The intratracheal bioavailability with respective to intravenous route was calculated by measuring the blood glucose reduction. The coarser particles of lactose in fractions of carrier containing a wide particle size distribution impacted in the preseperator of cascade impactor, and only the particle less than 10 microm size entered stage 0-stage 7. Formulation containing 1:1 mixture of Respitose ML006 (62%<50 microm) and Respitose ML003 (37.8%<50 microm) as carrier imparts well deaggregation of insulin, and higher deposition leads to 52.3% of fine particle fraction at 60 Lit/min and in vivo bioavailability of 82%. Insulin formulations containing 1:1 mixture of Respitose ML006 and Respitose ML003 as carrier can impart deeper deposition of drug particles and cause higher bioavailability. This suggests that carrier used in the formulation influenced the amount of insulin deposition in the alveolar region of the lung. Hence, it was concluded that the availability of insulin for systemic absorption depends on the particle size of the drug as well as the carrier lactose.


The effect of instrument variables on calculation factors for standardisation of colorimetric analyses.
A number of colorimetric methods, particularly enzyme activity assays, are usually standardised using calculation factors based on the molar absorptivity of a principle reactant or product. Such methods are subject to long-term variation. The relationship between long-term variation in results and instrument variables affecting calculation factors has not been quantitated. In this study, we have shown that, on a centrifugal analyser having a within-run coefficient of variation of less than 1%, instrument variables affecting calculation factor alone could result in changes in results of up to 8.5% over 75 days. We therefore advocate daily use of a solution of potassium dichromate to monitor instrument variables that can independently affect calculation factors and within-run imprecision. This procedure is useful for maintaining long-term performance and for differentiating problems of instrumental or chemical origin.


Monoclonal antibody produced against partially purified sporozoite antigen inhibits invasion of cells by sporozoites of avian Eimeria species.
Previous studies showed that molecules of in vitro-cultured primary turkey kidney cells bound to 23-, 40-, and 60- to 65-kDa antigens of sodium dodecyl sulfate-solubilized sporozoites of Eimeria adenoeides. Similar binding to antigens of three other species of avian Eimeria, E. tenella, E. acervulina, and E. meleagrimitis, is now reported. Strips containing the most avidly bound sporozoite antigen (approximately 40 kDa) were excised from the sodium dodecyl sulfate-polyacrylamide gels on which E. adenoeides antigens had been electrophoretically separated. The strips were homogenized and injected into mice to produce hybridoma cell lines. Twelve cell lines secreting monoclonal antibodies (McAb) that reacted with E. adenoeides sporozoites were detected. One of these McAb, H11C3, reacted with structures in the anterior tip of sporozoites of E. adenoeides and five other species of avian Eimeria. When included in the inoculation medium, this McAb significantly inhibited invasion of cultured kidney cells by sporozoites of E. adenoeides and E. tenella. In contrast, when the sporozoites were pretreated with McAb H11C3 and then washed free of the antibody, no inhibition of invasion was observed.


Non-digestible oligosaccharides scFOS/lcFOS facilitate safe subcutaneous immunotherapy for peanut allergy.
Improving the safety of subcutaneous immunotherapy (SCIT) for food allergy is necessary to reduce side effects and achieve long-term tolerance. We determined the effect of dietary supplementation with 1% non-digestible short- and long-chain fructo-oligosaccharides (scFOS/lcFOS) on safety and efficacy of SCIT using a peanut allergy mouse model. After sensitization, mice received a scFOS/lcFOS or control diet for the rest of the study. To study safety of SCIT, mice were dosed with a single subcutaneous injection of peanut extract (PE) or PBS. To study efficacy, mice were dosed subcutaneously (SCIT, 3 times/week) with PE or PBS for 3 weeks. Hereafter, acute allergic skin responses, anaphylactic shock symptoms and body temperature were assessed. To study the mechanism in vitro, the human IgE receptor (FcεRI)-transfected rat mast cell (RBL) line was sensitized with an oligoclonal pool of chimeric human (chu)IgE antibodies against bovine β-lactoglobulin (BLG) and incubated with the oligosaccharides before exposure to BLG to assess direct the effect on degranulation. scFOS/lcFOS reduced anaphylaxis caused by a single PE SCIT dose. scFOS/lcFOS alone also reduced the acute allergic skin response. Moreover, scFOS/lcFOS supplementation resulted in lower MMCP-1 levels in serum after PE SCIT dose compared to control diet, while antibody levels were not affected by the diet. In vitro incubation with scFOS/lcFOS at 0.5% suppressed the degranulation of IgE-sensitized RBL cells. However, dietary supplementation with scFOS/lcFOS did not improve the efficacy of SCIT. We show that scFOS/lcFOS diet improves the safety of SCIT, as evidenced by lower anaphylactic responses without compromising the efficacy in a mouse model for peanut allergy. This effect is likely to result from the suppression of mast cell effector function.


The quality of obstetrical clinical practice guidelines promulgated by a specialty society.
To review the methodologic quality of all obstetrical clinical practice guidelines developed by the Society of Obstetricians and Gynaecologists of Canada (SOGC) from 1992 to August 2001. Three reviewers independently assessed each of 37 guidelines according to a validated and reliable 37-item appraisal tool. A mean "global" score as well as three "dimension" scores were calculated for each guideline. The appraisal tool assessed each document according to three dimensions, which related to the rigour of its development (Dimension 1), its context and content (Dimension 2), and its application (Dimension 3). A repeated-measures analysis of variance was used to derive the interclass correlation coefficient. Mean global quality scores ranged from 8.1% to 54.0%. Only two guidelines were given a mean global score above 50% and 21 of the guidelines (56.8%) had mean global scores of less than 30%. Mean dimension scores were 19.9% for Dimension 1, relating to rigour of development, 47.3% for Dimension 2, relating to context and content, and 27.2% for Dimension 3, relating to application. The interclass correlation coefficient using a fixed-effects model was 0.72, reflecting reasonable agreement between the reviewers. Both the mean global scores and mean dimension-specific scores for the obstetrical clinical practice guidelines were lower than optimal. We were unable to reveal statistically significant improvements in guideline quality over time given the limited number of documents, but the scores for more recently drafted guidelines appear generally higher than earlier guidelines. This finding is consistent with new editorial policy adopted by the SOGC regarding guideline development. A broader review should be conducted by the SOGC on guidelines in development and under revision.