Characterization of two polyketide methyltransferases involved in the biosynthesis of the antitumor drug mithramycin by Streptomyces argillaceus.
A DNA chromosomal region of Streptomyces argillaceus ATCC 12596, the producer organism of the antitumor polyketide drug mithramycin, was cloned. Sequence analysis of this DNA region, located between four mithramycin glycosyltransferase genes, showed the presence of two genes (mtmMI and mtmMII) whose deduced products resembled S-adenosylmethionine-dependent methyltransferases. By independent insertional inactivation of both genes nonproducing mutants were generated that accumulated different mithramycin biosynthetic intermediates. The M3DeltaMI mutant (mtmMI-minus mutant) accumulated 4-demethylpremithramycinone (4-DPMC) which lacks the methyl groups at carbons 4 and 9. The M3DeltaM2 (mtmMII-minus mutant) accumulated 9-demethylpremithramycin A3 (9-DPMA3), premithramycin A1 (PMA1), and 7-demethylmithramycin, all of them containing the O-methyl group at C-4 and C-1', respectively, but lacking the methyl group at the aromatic position. Both genes were expressed in Streptomyces lividans TK21 under the control of the erythromycin resistance promoter (ermEp) of Saccharopolyspora erythraea. Cell-free extracts of these clones were precipitated with ammonium sulfate (90% saturation) and assayed for methylation activity using different mithramycin intermediates as substrates. Extracts of strains MJM1 (expressing the mtmMI gene) and MJM2 (expressing the mtmMII gene) catalyzed efficient transfer of tritium from [(3)H]S-adenosylmethionine into 4-DPMC and 9-DPMA3, respectively, being unable to methylate other intermediates at a detectable level. These results demonstrate that the mtmMI and mtmMII genes code for two S-adenosylmethionine-dependent methyltransferases responsible for the 4-O-methylation and 9-C-methylation steps of the biosynthetic precursors 4-DPMC and 9-DPMA3, respectively, of the antitumor drug mithramycin. A pathway is proposed for the last steps in the biosynthesis of mithramycin involving these methylation events.


[ Food allergy ].
Food allergies' following food incompatibilities, which are not caused immunologically. Mostly allergic symptoms are caused by cow's milk or chicken eggs. Allergic reactions are preceded by sensitizing events; certain characteristics of foodstuffs and conditions in the human body facilitate their development. Gastrointestinal symptoms very often are just accompanying signs. In differential diagnosis the so-called "pseudo-allergies' following food ingestion have to be separated. Most important diagnostic measures are clinical history, prick-/scratch test, RAST, gastrointestinal provocation and abstinence test. The therapeutic program consists of allergen abstinence, avoiding all allergy-arousing factors, oral desensitizing and pharmaceutical treatment.


Anti-TGF-beta antibody blocks enamel matrix derivative-induced upregulation of p21WAF1/cip1 and prevents its inhibition of human oral epithelial cell proliferation.
We have previously demonstrated that porcine enamel matrix derivative (EMD) contains TGF-beta 1 (or a TGF-beta-like substance), and that EMD rapidly translocates smad2, which is an effector of the TGF-beta signaling pathway, into the nucleus and modulates the proliferation of both human gingival fibroblastic and oral epithelial cells in a cell type-specific manner. To investigate the involvement of TGF-beta in the growth modulatory action of EMD, two approaches have been used in the present study: i) a neutralizing anti-TGF-beta antibody to block EMD action, and ii) authentic porcine TGF-beta 1 to compare with EMD. Both in epithelial and fibroblastic cells, TGF-beta 1 closely mimicked EMD in nuclear accumulation of smad2, phosphorylation of MAP kinase family members, and consequent cell type-specific growth modulation. Anti-TGF-beta antibody, at levels which completely blocked TGF-beta 1-induced smad2 translocation, strongly blocked EMD-induced smad2 translocation. This antibody also blocked other actions of EMD in epithelial cells, i.e. p38-MAP kinase (p38-K) phosphorylation, p21WAF1/cip1 expression, and inhibition of DNA synthesis. In support of our previous proposal, these data suggest that TGF-beta 1 (or a TGF-beta-like substance), which is delivered as a principal bioactive factor in EMD, inhibits epithelial cell proliferation probably by a smad2-mediated, p21WAF1/cip1-dependent mechanism. However, the same neutralizing antibody failed to convincingly block EMD-induced fibroblastic proliferation, which suggests that EMD may contain additional unidentified mitogenic factor(s), which act in combination with TGF-beta to fully stimulate fibroblastic proliferation.


Cost-effectiveness of elbasvir/grazoprevir use in treatment-naive and treatment-experienced patients with hepatitis C virus genotype 1 infection and chronic kidney disease in the United States.
Among patients with chronic kidney disease (CKD) in the United States, HCV infection causes significant morbidity and mortality and results in substantial healthcare costs. A once-daily oral regimen of elbasvir/grazoprevir (EBR/GZR) for 12 weeks was found to be a safe and efficacious treatment for HCV in patients with CKD. We evaluated the cost-effectiveness of EBR/GZR in treatment-naïve and treatment-experienced CKD patients compared with no treatment (NoTx) and pegylated interferon plus ribavirin (peg-IFN/RBV) using a computer-based model of the natural history of chronic HCV genotype 1 infection, CKD and liver disease. Data on baseline characteristics of the simulated patients were obtained from NHANES, 2000-2010. Model inputs were estimated from published studies. Cost of treatment with EBR/GZR and peg-INF/RBV were based on wholesale acquisition cost. All costs were from a third-party payer perspective and were expressed in 2015 U.S. dollars. We estimated lifetime incidence of liver-related complications, liver transplantation, kidney transplantation, end-stage live disease mortality and end-stage renal disease mortality; lifetime quality-adjusted life years (QALY); and incremental cost-utility ratios (ICUR). The model predicted that EBR/GZR will significantly reduce the incidence of liver-related complications and prolong life in patients with chronic HCV genotype 1 infection and CKD compared with NoTx or use of peg-IFN/RBV. EBR/GZR-based regimens resulted in higher average remaining QALYs and higher costs (11.5716, $191 242) compared with NoTx (8.9199, $156 236) or peg-INF/RBV (10.2857, $186 701). Peg-IFN/RBV is not cost-effective, and the ICUR of EBR/GZR compared with NoTx was $13 200/QALY. Treatment of a patient on haemodialysis with EBR/GZR resulted in a higher ICUR ($217 000/QALY). Assuming a threshold of $100 000 per QALY gained for cost-effectiveness, use of elbasvir/grazoprevir to treat an average patient with CKD can be considered cost-effective in the United States.


Reduction of dietary protein and phosphorus in the Modification of Diet in Renal Disease Feasibility Study. The MDRD Study Group.
The Modification of Diet in Renal Disease (MDRD) Feasibility Study was designed to test procedures and evaluate the feasibility of a full-scale clinical trial aimed at assessing the effects of reduction of dietary protein and phosphorus on progression of renal disease. Ninety-six patients with chronic renal insufficiency were randomly assigned to different diets in one of two studies depending on their glomerular filtration rate. The diets contained three different protein and phosphorus levels: moderate diet = 1.3 g protein per kilogram per day and 16 to 20 mg phosphorus per kilogram per day; low diet = 0.575 g protein per kilogram per day and 5 to 10 mg phosphorus per kilogram per day; and very low diet with keto or amino acids = 0.28 g protein per kilogram per day and 4 to 9 mg phosphorus per kilogram per day. Eight-five patients were monitored for at least 6 months; maximum follow-up was 22 months. Compliance with study diets was measured monthly using urea nitrogen appearance and 3-day diet diaries plus one 24-hour recall. The main outcome measure was change or maintenance of glomerular filtration rate. Data were analyzed by analysis of variance and paired t tests. Mean dietary protein intake, as determined by urea nitrogen appearance, decreased significantly in participants assigned to the diets low and very low in protein and phosphorus (P < .05). Overall, the follow-up protein intake (based on urea nitrogen appearance) as a percentage of baseline ranged from 45.8% to 83.1%. Analysis of diet diaries showed better dietary adherence than indicated by urea nitrogen appearance. Review of the exchange methodology used in dietary instruction suggests that imprecision of the exchange lists may have been a factor in the difficulty study participants had in achieving +/- 10% of the target protein goal. Based on the MDRD Feasibility Study, the protocol for the full-scale study was modified to include protein counting instead of food exchange methodology to monitor protein intake.


Ionization and fragmentation of isomeric van der Waals complexes embedded in helium nanodroplets.
The ionization and charge transfer processes, which occur when a doped helium droplet undergoes electron impact, are studied for droplets doped with van der Waals complexes with various structures and electrostatic moments. The mass spectra of the two isomers of hydrogen cyanide complexed with either cyanoacetylene or acetylene in helium droplets were obtained using optically selected mass spectrometry, and show that the structure of the complex has a large effect on the fragmentation pattern. The resulting fragmentation pattern is consistent with an ionization process in which charge steering strongly influences the site of initial ionization. The observed dissociation products may also be subject to caging by the helium matrix.


The mental health crisis.
As public funding drops and need soars, hospitals are working with other providers to find solutions to this problem.


Brain Metastasis and Renal Cell Carcinoma: Prognostic Scores Assessment in the Era of Targeted Therapies.
This study aimed at exploring several brain metastatic prognostic scores in patients with renal cell carcinoma. We retrospectively analyzed data of 93 metastatic renal cell carcinoma patients who were diagnosed with brain metastases between October 2005 and July 2016 who received targeted therapy. Potential prognostic factors (RTOG RPA, BS-BM, and a newly developed score CERENAL) were analyzed. A total of 75 patients received targeted therapy. All scores showed prognostic value in progression-free survival after first-line treatment with CERENAL being the sole independent prognostic factor associated with improved duration of first-line treatment. Both RTOG RPA and CERENAL were potential prognosticators for overall survival, whereas only the CERENAL score was associated with prolonged disease-specific survival. Several prognostic scores can be useful to predict survival of patients with brain metastases from renal cancer, especially the newly developed CERENAL score.


Shp-2 positively regulates brain-derived neurotrophic factor-promoted survival of cultured ventral mesencephalic dopaminergic neurons through a brain immunoglobulin-like molecule with tyrosine-based activation motifs/Shp substrate-1.
To examine the roles of Shp-2, a cytoplasmic tyrosine phosphatase, in neuronal survival, we generated and used recombinant adenoviruses expressing wild type and phosphatase-inactive (C/S), phosphatase domain-deficient (delta P) and constitutively active (D61A and E76A) mutants of Shp-2. We found that wild-type Shp-2 enhanced brain-derived neurotrophic factor (BDNF)-promoted survival of cultured ventral mesencephalic dopaminergic neurons. In contrast, the C/S and delta P mutants of Shp-2 did not affect survival. In addition, the constitutively active D61A and E76A mutants mimicked BDNF and promoted survival. Furthermore, to examine the effects of BIT/SHPS-1, a substrate of Shp-2, on the BDNF-promoted survival, we generated adenovirus vectors expressing wild-type BIT/SHPS-1 and its 4F mutant in which all tyrosine residues in the cytoplasmic domain of BIT/SHPS-1 were replaced with phenylalanine. We found that BDNF-promoted survival of cultured mesencephalic dopaminergic neurons was enhanced by expression of the 4F mutant but not of wild-type BIT/SHPS-1. In addition, we found that co-expression of wild-type BIT/SHPS-1 with Shp-2 significantly enhanced the survival-promoting effect of BDNF on cultured mesencephalic dopaminergic neurons. These results indicated that Shp-2 positively regulates the survival-promoting effect of BDNF on cultured ventral mesencephalic dopaminergic neurons. Dephosphorylation of BIT/SHPS-1 by Shp-2 may participate in BDNF-stimulated survival signaling.


Relationship between drug resistance and HIV-1 disease progression or death in patients undergoing resistance testing.
To evaluate factors associated with drug resistance detected by genotypic antiretroviral resistance testing (GART), and to determine the association between the level of resistance and subsequent human immunodeficiency type 1 (HIV-1) disease progression or death. Observational cohort study. We identified highly active antiretroviral therapy (HAART)-treated patients who had GART as part of clinical management. Factors associated with greater numbers of resistance mutations were assessed by ordinal logistic regression. Survival analysis was used to assess time to a new opportunistic condition or death following GART. A total of 572 patients were identified who had GART: of these, 50% had 0-2 resistance mutations, 33% had 3-6 mutations, and 17% had >/= 7 mutations. In multivariate analysis, prolonged use of HAART in the setting of incomplete viral suppression was significantly associated with more drug resistance. Patients with fewer resistance mutations were significantly more likely to achieve viral suppression after GART than patients with more mutations. Compared to patients with two or less resistance mutations, those with three to six mutations, or seven or more mutations were not at higher risk of HIV-1 disease progression or death over a median follow-up of 15 months. In contrast, continued HAART use following GART was strongly associated with slower disease progression, particularly at lower CD4 cell counts. These results support the hypothesis the drug-resistant HIV-1 may be less pathogenic than wild-type virus, and that continued use of HAART might provide clinical benefit, despite persistent viremia and HIV-1 drug resistance.


Fine-needle aspiration of metastatic radiation-induced cutaneous epithelioid angiosarcoma of the breast to the liver: A diagnostic dilemma.
Radiation therapy is a significant risk factor for the development of angiosarcoma. With the increase in breast conservation treatment, the incidence of cutaneous radiation-induced angiosarcoma of the breast is rising. If the angiosarcoma demonstrates epithelioid features, the tumor cells may present a diagnostic dilemma on fine-needle aspiration cytology. We present a case of metastatic radiation-induced cutaneous epithelioid angiosarcoma of the breast to the liver and a review of the literature.


The anal sphincter in patients with myotonic muscular dystrophy.
The objective of this prospective study was to determine anal sphincter function and thickness of the anal musculature in patients with myotonic muscular dystrophy. Manometric studies were performed in 16 patients with myotonic dystrophy and in 16 healthy controls. Patients had significantly lower basal and squeeze pressures than control subjects (P less than 0.01). The results of ultrasonographic studies of the anal canal in 7 patients and 7 control subjects suggest that this decrease in muscle strength is partly explained by muscular atrophy. In addition, patients with myotonic dystrophy showed exaggerated rebound contractions following and sphincter relaxation that was induced by rectal distention. The pattern of this response and the results of electromyographic studies in 6 patients with myotonic dystrophy suggest that such abnormalities are explained by a neurogenic defect rather than a myotonic response of the anal musculature. It is concluded that patients with myotonic dystrophy show a multitude of defects in the anal sphincter that are an expression of myopathy, muscular atrophy, and neural abnormalities.


Intravascular site-specific delivery of a therapeutic antisense for the inhibition of restenosis.
Polymeric nanoparticles (NPs) have been implicated as potential gene carriers in the treatment of various genetic and acquired diseases. In this work we investigated the efficacy of NPs as gene carrier for intravascular gene therapy in animal models of restenosis. A therapeutic antisense against the monocyte chemotactic protein-1 (anti-MCP-1) was encapsulated into the poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) using double-emulsion/solvent evaporation technology. Laser defractometer was used to assess size distribution of NPs. Particle morphology was assessed by scanning electron microscopy (SEM). DNA content in NPs was determined by DNA extraction using TE buffer from a chloroform solution dissolved a known amount of NPs. DNA concentration was assayed by spectrophotometer. In vitro DNA release was performed in the TE buffer at 37 degrees C utilizing double-chamber diffusion cell. NPs loaded with pEGFP and anti-MCP-1 gene were tested in SMC cell culture for transduction efficiency. The anti-MCP-1 NPs were further evaluated in rabbit vein grafting and carotid artery injury models for their potential in inhibition of restenosis. The NPs demonstrated a steady in vitro release of DNA with approximately 95% of total enclosed DNA released within 30 days. Anti-sense MCP-1 expression was confirmed in arterial tissues with single infusion of the therapeutic NPs into the injured rabbit carotid arteries. The intima/media ratio of arteries treated with the anti-MCP-1 NP was reduced by 43% compared with control groups following a 2-week treatment. In a rabbit jugular vein-to-artery-bypass grafting model, animals with local-infusion of anti-MCP-1 NPs also demonstrated a significantly lower intimal hyperplasia than that of control groups with no or free antisense treatment. Collectively, our data revealed that local delivered anti-MCP-1 NPs effectively inhibited experimental restenosis.


Microwave photochemical reactor for the online oxidative decomposition of p-hydroxymercurybenzoate (pHMB)-tagged proteins and their determination by cold vapor generation-atomic fluorescence detection.
A novel method is presented for the characterization and determination of thiolic proteins. After the labeling with p-hydroxymercurybenzoate, the pHMB-labeled proteins underwent on-line oxidation with a novel microwave (MW)/UV photochemical reactor, followed by cold vapor generation-atomic fluorescence spectrometry (CVG-AFS) detection. The MW/UV process led to the conversion of pHMB to Hg(II) with a yield of 89.0 ± 0.5% without using chemical oxidizing reagents and avoiding the use of toxic carcinogenic compounds. Hg(II) was reduced to Hg(0) in a knotted reaction coil with NaBH4 solution, stripped from the solution by an argon flow and detected. The chromatographic method for labeled thiolic peptides was linear in the 0.2-100 μmol L(-1) range, with a LOD as mercury of 57 nmol L(-1). This system has proven to be a useful interface for liquid chromatography coupled with CVG-AFS in the determination and characterization of thiolic proteins. This method has been applied to the determination of thiolic peptides after tryptic digestion of serum albumins from different species (human, bovine, rat, horse, and sheep).


Adult Schistosoma mansoni express cathepsin L proteinase activity.
This report presents the deduced amino acid sequence of a novel cathepsin L proteinase from Schistosoma mansoni, and describes cathepsin L-like activity in extracts of adult schistosomes. Using consensus primers specific for cysteine proteinases, gene fragments were amplified from adult S. mansoni cDNA by PCR and cloned. One of these fragments showed marked identity to Sm31, the cathepsin B cysteine proteinase of adult S. mansoni, whereas another differed from Sm31 and was employed as a probe to isolate two cDNAs from an adult S. mansoni gene library. Together these cDNAs encoded a novel preprocathepsin L of 319 amino acids; this zymogen is predicted to be processed in vivo into a mature, active cathepsin L proteinase of 215 amino acids. Closest homologies were with cathepsins L from rat, mouse, and chicken (46-47% identity). Southern hybridization analysis suggested that only one or a few copies of the gene was present per genome, demonstrated that its locus was distinct from that of Sm31, and that a homologous sequence was present in Schistosoma japonicum. Because these results indicated that schistosomes expressed a cathepsin L proteinase, extracts of adult S. mansoni were examined for acidic, cysteine proteinase activity. Based on rates of cleavage of peptidyl substrates employed to discriminate between classes of cysteine proteinases, namely cathepsin L (Z-phe-arg-AMC), cathepsin B (Z-arg-arg-AMC) and cathepsin H (Bz-arg-AMC), the extracts were found to contain vigorous cathepsin L-like activity.(ABSTRACT TRUNCATED AT 250 WORDS)


Aggressive uterine sarcoma with rhabdoid features: diagnosis by peritoneal fluid cytology and absence of INI1 gene mutation.
We report a primary uterine sarcoma with classic histologic, immunohistochemical, and ultrastructural features of a malignant extrarenal rhabdoid tumor (MERT). It arose in a 71-year-old woman who presented with postmenopausal bleeding, ascites, and a right pelvic mass. Malignant cells with rhabdoid morphology were identified by cytologic examination of the peritoneal fluid. Exploratory laparotomy revealed a 10-cm right adnexal mass and disseminated peritoneal tumor. Pathologic study showed diffuse expansion of the endometrial stroma by rhabdoid-like cells with transmural infiltration of the myometrium and extensive involvement of uterine serosa and right ovary by tumor. Neoplastic cells were immunoreactive for vimentin, cytokeratin, and epithelial membrane antigen, and cytoplasmic whorls of intermediate filaments were observed by electron microscopy. Fluorescence in situ hybridization (FISH) studies with chromosome 22-specific probes showed no loss of the INI1 gene, and no coding sequence mutation was identified.


Effect of FKBP65, a putative elastin chaperone, on the coacervation of tropoelastin in vitro.
FKBP65 is a protein of the endoplasmic reticulum that is relatively abundant in elastin-producing cells and is associated with tropoelastin in the secretory pathway. To test an earlier suggestion by Davis and co-workers that FKBP65 could act as an intracellular chaperone for elastin, we obtained recombinant FKBP65 (rFKBP65) by expressing it in E. coli and examined its effect on the coacervation characteristics of chicken aorta tropoelastin (TE) using an in vitro turbidimetric assay. Our results reveal that rFKBP65 markedly promotes the initiation of coacervation of TE without significantly affecting the temperature of onset of coacervation. This effect shows saturation at a 1:2 molar ratio of TE to rFKBP65. By contrast, FKBP12, a peptidyl prolyl isomerase, has a negligible effect on TE coacervation. Moreover, the effect of rFKBP65 on TE coacervation is unaffected by the addition of rapamycin, an inhibitor of peptidyl prolyl isomerase (PPIase) activity. These observations rule out the involvement of the PPIase activity of rFKBP65 in modulating the coacervation of TE. Additional experiments using a polypeptide model of TE showed that rFKBP65, while promoting coacervation, may retard the maturation of this model polypeptide into larger aggregates. Based on these results, we suggest that FKBP65 may act as an elastin chaperone in vivo by controlling both the coacervation and the maturation stages of its self-assembly into fibrils.


Physician workforce planning and the transition to primary health care in former socialist countries.
With the collapse of the Soviet Union, countries in Eastern Europe and the Newly Independent States inherited a physician workforce that was often too large, dominated by specialists, and poorly prepared for the transition to primary health care and the addition of the family/general practice specialty. We examine attempts in selected countries to plan the future physician workforce, while attempting to reduce the size of the workforce and train physicians to lead the transition to primary health care (PHC). We look the impact these efforts have had on the current workforce and will have on the future physician workforce. With few exceptions, the first move after independence was to reduce the inputs into the physician workforce in an attempt to reduce the size of the workforce, considered large by western standards, in 1990 between 350 and 400 per 100, 000 population compared to the EU average of 299. These reductions often did not result from planning and ignored the lengthy physician training process, leading to concerns for the future supply of physicians and the conclusion that many other factors were influencing the number of physicians. At the same time, two methods were being employed to rapidly prepare physicians for PHC, retraining of existing physicians for the short-term and the establishment of training programs in the faculties of medicine to train family/general practitioners (GPs) for the long-term. GPs per 100,000 population remained at about 102 throughout the period in the original EU countries, but in the new EU countries went from 51 in 1991 to 63 in 2002. The success of the programs was varied and often depended on the overall organization of the physician workforce, the status of the new family physician within the workforce and the commitment at the national level to the transition to PHC. After over a decade of independence, there is still a struggle to have a physician workforce with the right numbers, the right specialty mix, and practicing in the right locations.


[Hygienic evaluation of pollution in the area of a chemically-dangerous enterprise].
Pollution was hygienically evaluated in the area of the former chemically dangerous OAO "Kaprolaktam-Dzerzhinsk" enterprise. The soils around its production buildings were the test subject. Sampling, chemical analysis, and hygienic evaluation of the results of the analysis were made. The work done allowed the authors to plot pollution by both area and depth. The findings may be used to choose a decontamination procedure, by taking into account the degree of pollution.


Interactions between the rostral ventral medulla and other central sites involved in vasomotor regulation.
We showed earlier that the rostral ventral medulla can be functionally and anatomically divided into the rostral ventrolateral medulla (RVLM) and the rostral ventromedial medulla (RVMM). In this study, we examined the relationship between the lateral hypothalamus and these two medullary sites. Electrical stimulation of the lateral hypothalamus produced a pressor response mediated by increases in renal and mesenteric vascular resistance. Inactivating RVLM had no effect on this response whereas inactivating RVMM significantly blunted the pressor response and the increases in renal and mesenteric resistance. In other studies, inactivating dorsomedullary sites significantly reduced arterial pressure. Unlike RVLM, these depressor responses were not significantly altered by reducing tidal volume. Inactivating sites 0.5 mm medial and lateral to the RVMM, as well as a site 0.5 mm caudal to the RVLM, resulted in depressor responses that were unaffected by reducing tidal volume. However, inactivation of a site 1.0 mm caudal to the RVLM but still within the ventrolateral medullary pressor area, resulted in a depressor response that was enhanced by reduced tidal volume. Together, these data demonstrate a functional differentiation of medullary sites controlling vasomotor outflow.


Copper availability and accumulation by Portulaca oleracea Linn. stem cutting.
The Cu accumulation capacity of stem cuttings of Portulaca oleracea was assessed in two different soils, alfisol and vertisol, which differ from each other mainly due to their clay content - several fold lower in the former than in the latter. The DTPA extractable concentration of Cu in soil and the level of its accumulation in stem cuttings were, therefore, greater in alfisol than in vertisol. In the mixed soils, the DTPA extractable as well as plant accumulated levels of Cu decreased with an increase in the proportion of vertisol. With an increase in the soil available and plant accumulated Cu, a decrease has been observed in the ability of stem cuttings to regenerate leaves and roots. As a consequence biomass reduction has been observed with an increase in the concentration of Cu in the soil and plant body. In view of comparatively low availability of Cu in vertisol, regeneration of stem cuttings occurred up to a concentration of 1600 microg of Cu/g of soil, whereas in alfisol it occurred only up to 200 microg of Cu/g of soil. The results of the present study showed that P. oleracea stem cutting is an accumulator of Cu and the availability of Cu in soils could be manipulated through addition of clay since the plant available concentration of Cu is less in the soil (vertisol) with high clay content.


Reprint of: Comparison of affinity tags for protein purification.
Affinity tags are highly efficient tools for purifying proteins from crude extracts. To facilitate the selection of affinity tags for purification projects, we have compared the efficiency of eight elutable affinity tags to purify proteins from Escherichia coli, yeast, Drosophila, and HeLa extracts. Our results show that the HIS, CBP, CYD (covalent yet dissociable NorpD peptide), Strep II, FLAG, HPC (heavy chain of protein C) peptide tags, and the GST and MBP protein fusion tag systems differ substantially in purity, yield, and cost. We find that the HIS tag provides good yields of tagged protein from inexpensive, high capacity resins but with only moderate purity from E. coli extracts and relatively poor purification from yeast, Drosophila, and HeLa extracts. The CBP tag produced moderate purity protein from E. coli, yeast, and Drosophila extracts, but better purity from HeLa extracts. Epitope-based tags such as FLAG and HPC produced the highest purity protein for all extracts but require expensive, low capacity resin. Our results suggest that the Strep II tag may provide an acceptable compromise of excellent purification with good yields at a moderate cost.


Significant elements of community involvement in participatory action research: evidence from a community project.
Participatory action research (PAR) has been heralded as an important research methodology to address issues of research relevance, community involvement, democracy, emancipation and liberation. Increasingly, nurse researchers are turning to PAR as a method of choice. Although nursing interest in PAR is expanding little is known about how to successfully involve the community in research. This article attends to this dearth of information by presenting the results of a study investigating the significant elements of community involvement in PAR. Through the use of qualitative research methods, five themes emerged that describe the community participation process: (a) planning for participation, (b) the structural components of community participation, (c) living the philosophy, (d) enhancing the credibility, and (e) the type of leadership required to facilitate community participation. It is hoped that by sharing these results others may consider the knowledge gleaned from this project as they plan and proceed with the challenges and rewards inherent in PAR.


The association between vaginal bacterial composition and miscarriage: a nested case-control study.
To characterise vaginal bacterial composition in early pregnancy and investigate its relationship with first and second trimester miscarriages. Nested case-control study. Queen Charlotte's and Chelsea Hospital, Imperial College Healthcare NHS Trust, London. 161 pregnancies: 64 resulting in first trimester miscarriage, 14 in second trimester miscarriage and 83 term pregnancies. Prospective profiling and comparison of vaginal bacteria composition using 16S rRNA gene-based metataxonomics from 5 weeks' gestation in pregnancies ending in miscarriage or uncomplicated term deliveries matched for age, gestation and body mass index. Relative vaginal bacteria abundance, diversity and richness. Pregnancy outcomes defined as first or second trimester miscarriage, or uncomplicated term delivery. First trimester miscarriage associated with reduced prevalence of Lactobacillus spp.-dominated vaginal microbiota classified using hierarchical clustering analysis (65.6 versus 87.7%; P = 0.005), higher alpha diversity (mean Inverse Simpson Index 2.5 [95% confidence interval 1.8-3.0] versus 1.5 [1.3-1.7], P = 0.003) and higher richness 25.1 (18.5-31.7) versus 16.7 (13.4-20), P = 0.017), compared with viable pregnancies. This was independent of vaginal bleeding and observable before first trimester miscarriage diagnosis (P = 0.015). Incomplete/complete miscarriage associated with higher proportions of Lactobacillus spp.-depleted communities compared with missed miscarriage. Early pregnancy vaginal bacterial stability was similar between miscarriage and term pregnancies. These findings associate the bacterial component of vaginal microbiota with first trimester miscarriage and indicate suboptimal community composition is established in early pregnancy. While further studies are required to elucidate the mechanism, vaginal bacterial composition may represent a modifiable risk factor for first trimester miscarriage. Vaginal bacterial composition in first trimester miscarriage is associated with reduced Lactobacillus spp. abundance and is independent of vaginal bleeding.


Pathways of care the first year after moderate and severe traumatic brain injury-discharge destinations and outpatient follow-up.
To determine the pathways of care within the first year after traumatic brain injury (TBI) and to explore whether provided care is related to residual impairments. Retrospective study of 343 patients with moderate and severe TBI admitted to a Level-1 trauma centre. Discharge destinations from hospital to home and frequency of outpatient visits were determined. Outcome was defined 1 year after injury by the Extended Glasgow Outcome Scale and Return to Work. Most (94%) patients had returned home 1-year after injury despite cognitive (76%), behavioural (67%) and physical (55%) impairments. One in four patients was severely disabled and 32% had resumed work on a previous level. Two-thirds of all patients went home as secondary discharge destination and 50% needed inpatient rehabilitation. Almost half of patients needed outpatient care, mostly for behavioural and cognitive impairments. One in 10 patients consulted a psychiatrist, with 55% unfavourable outcome. Of those patients initially discharged to home without follow-up, eventually 10% needed outpatient rehabilitation. One-year after injury most patients had returned home with residual impairments and frequent medical consultations. This finding warrants further investigation to define appropriate aftercare by various medical specialists aimed at long-term community integration.


[Evaluation of external respiratory function in patients with chronic obstructive bronchitis using radionuclide methods].
A trial was designed to study shifts in various mechanisms of external respiration (ER) distress in chronic obstructive bronchitis by type of respiration insufficiency (RI). Combined clinicoroentgenological, spirographic, endoscopic and radionuclide (133Xe radiopneumography and 99Tc scintigraphy) examinations were conducted in 66 patients (41 males and 25 females aged 20-60 with 2-17-year history of chronic obstructive bronchitis) versus 30 healthy controls. ER parameters assessed in 4 groups of patients (with-out resting RI, with obstructive and mixed RI variants) by+radiopneugraphy and scintigraphy were found abnormal for either lung and both lungs as a whole. The abnormalities recorded correlated with RI variant. The following parameters appeared most informative in studying ER in the bronchitis patients with radionuclide techniques: the volume of ventilated alveoli, respiratory capacity, total and functional residual lung capacity, index ventilation/blood flow, capillary blood flow, time of half elimination of Xe from the alveoli and vascular bed. The above parameters change for the worse with progressive deterioration of external respiration.


Nephrotic syndrome: new concepts in the pathophysiology of sodium retention.
The nephrotic syndrome is a frequent clinical condition characterized by fluid and salt retention. Although several theories have been put forward to explain the salt-retaining status, recent data have confirmed previous renal micropuncture observations indicating that the distal nephron is the site for increased salt reabsorption, eventually leading to sodium retention. Target proteomic approaches and immunocytochemistry experiments have identified the epithelial sodium channel (ENaC) and basolateral Na+,K+-ATPase as the main transport proteins responsible for increased transepithelial sodium reabsorption in various forms of experimental nephrotic syndrome. Although the fine-tuning for the up-regulation of these transporters has not been so far elucidated, it is clear from clinical studies that the use of amiloride, a selective, dose-dependent ENaC inhibitor, is an appropriate tool to reduce distal sodium reabsorption and thus to offset edema formation.


Predicting atelectasis by assessment of collateral ventilation prior to endobronchial lung volume reduction: a feasibility study.
The advent of endoscopic lung volume reduction (ELVR), especially relying on valve technology to achieve atelectasis, has aroused new interest in the assessment of collateral ventilation, which has been implicated in ELVR failures. We are reporting on the use of a catheter-based device that measures airway pressures and flows, and calculates airway resistance in vivo. To assess the safety of this catheter-based system and the feasibility of obtaining measurements predictive of atelectasis after ELVR. Patients undergoing ELVR were prospectively included in this double-blind cohort study. Each lobe targeted for ELVR was blocked with a catheter system (Chartis® System; Pulmonx, Inc., Redwood, Calif., USA); pressures and flows were assessed continuously. The primary endpoints were to evaluate the safety and feasibility; the secondary endpoint was to assess whether there was a relationship between the measurements and the incidence of atelectasis following ELVR. From June 2008 to November 2008, 25 patients were included in the study. All procedures could be performed without any complications. Due to pneumothorax in 1 case and inability to assess the catheter-based measurements in 4 cases, the final analysis included 20 patients. Atelectasis occurred in 8 out of 20 cases following implantation. In 18 patients (90%), the resistance measurements correlated with the postimplantation atelectasis visualized on a chest X-ray; in 2 patients (10%), a mismatch was detected. Resistance measurements were safely and successfully achieved. In 90% of the analyzable cases, the resistance measurements correlated with the occurrence of atelectasis after ELVR. The clinical impact of these findings will need to be evaluated in subsequent trials.


Thermally driven smoothening of molecular thin films: Structural transitions in n-alkane layers studied in real-time.
We use thermal annealing to improve smoothness and to increase the lateral size of crystalline islands of n-tetratetracontane (TTC, C44H90) films. With in situ x-ray diffraction, we find an optimum temperature range leading to improved texture and crystallinity while avoiding an irreversible phase transition that reduces crystallinity again. We employ real-time optical phase contrast microscopy with sub-nm height resolution to track the diffusion of TTC across monomolecular step edges which causes the unusual smoothing of a molecular thin film during annealing. We show that the lateral island sizes increase by more than one order of magnitude from 0.5 μm to 10 μm. This desirable behavior of 2d-Ostwald ripening and smoothing is in contrast to many other organic molecular films where annealing leads to dewetting, roughening, and a pronounced 3d morphology. We rationalize the smoothing behavior with the highly anisotropic attachment energies and low surface energies for TTC. The results are technically relevant for the use of TTC as passivation layer and as gate dielectric in organic field effect transistors.


Nursing Theory in Virtual Care.
Individuals are increasingly seeking care in virtual care environments as the healthcare system evolves. Nurses are well poised to enhance their scope of practice into these environments that can transcend time, space, and location. The author discusses both how virtual care environments are seen in today's healthcare systems and how nursing theory and research will contribute to enhancing quality care for individuals in virtual care environments.


[Contribution of in vivo confocal microscopy to limbal tumor evaluation].
To explore tumors of the limbus with a new in vivo confocal microscope and to compare the images to histology results. We evaluated three tumors in three patients with the Heidelberg Retina Tomograph II, Rostock Cornea Module. A diagnostic and therapeutic excision with adjunctive cryotherapy was performed for each individual. Confocal microscopy was compared to histopathologic sections. Histology identified two dysplasias and one carcinoma in situ. The main pathological features were visible on our images: cytonuclear atypias, epithelial folds into an inflammatory and vascularized conjunctival stroma, fine vessels perpendicular to the surface, a clear limit with normal epithelium, papillomatous organization, and abnormal keratinization. Our preliminary study showed that this type of limbal tumor could be explored using in vivo confocal microscopy. We were not able to determine whether there was a microinvasion. This new method could be a diagnostic aid, especially for atypical lesions and for follow-up because of frequent recurrences. Other studies are necessary to confirm our hypothesis.


Pegfilgrastim: current and future perspectives in the treatment of chemotherapy-induced neutropenia.
Myeloid colony-stimulating factors (granulocyte colony-stimulating factor [G-CSF] and granulocyte-macrophage colony-stimulating factor) are commonly used in clinical practice for the prevention of anticancer chemotherapy-induced neutropenia and its potentially life-threatening complications. Pegfilgrastim is a novel recombinant human G-CSF pharmaceutically developed by covalent binding of a polyethylene glycol molecule to the N-terminal sequence of filgrastim. Due to its unique neutrophil-mediated clearance, pegfilgrastim can be administered once per chemotherapy cycle. Clinical trials have demonstrated that a single, fixed, subcutaneous dose of pegfilgrastim is comparable in safety and efficacy to daily injections of filgrastim for decreasing the incidence of infection following myelosuppressive chemotherapy in patients with cancer. Recent trials have been conducted to evaluate the use of pegfilgrastim in different clinical settings, including support of dose-dense regimens, mobilization and transplantation of hematopoietic stem cells.


Natural history of celiac disease autoimmunity in a USA cohort followed since 1974.
The natural history and the possible changes of celiac disease (CD) prevalence over time are still unclear. 1) To establish whether loss of tolerance to gluten may occur at any age; 2) to investigate possible changes of CD prevalence over time; and 3) to investigate CD-related co-morbidities. We analyzed 3,511 subjects with matched samples from 1974 (CLUE I) and 1989 (CLUE II). To avoid a selection bias regarding survival, we also screened 840 CLUE I participants who deceased after the 1974 survey. Outcome measure. CD autoimmunity (positivity to auto-antibodies) over time. CD autoimmunity was detected in seven subjects in 1974 (prevalence 1:501) and in an additional nine subjects in 1989 (prevalence 1:219). Two cases of CD autoimmunity were found among the 840 subjects deceased after CLUE I. Compared to controls, untreated CD subjects showed increased incidence of osteoporosis and associated autoimmune disorders, but they did not reach statistical significance. During a 15-year period CD prevalence increased 2-fold in the CLUE cohort and 5-fold overall in the US since 1974. The CLUE study demonstrated that this increase was due to an increasing number of subjects that lost the immunological tolerance to gluten in their adulthood.


Expression of Ca2+ -activated K+ channels in human dermal fibroblasts and their roles in apoptosis.
Cell proliferation and apoptosis are responsible for maintaining normal tissue homeostasis, and K(+) currents play important roles in regulating the physiological balance between them. This function of Ca(2+)-activated K(+) (K(Ca)) channels has been demonstrated in many types of tissues, but not in dermal fibroblasts. We investigated the expression of K(Ca) channels and their effects on proliferation and apoptosis in human dermal fibroblasts. We used discoidin domain receptor 2 immunostaining to identify human dermal fibroblasts, and reverse transcription polymerase chain reaction, Western blot analysis and electrophysiological patch clamp recordings to evaluate the expression and characteristics of 3 members of the K(Ca) channel family, large-conductance K(Ca) (BK), intermediate-conductance K(Ca) (IK) and small-conductance K(Ca) channels. We also used the 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, Hoechst 33258 staining and Depsipher staining to investigate the effects of K(Ca) channels on cell proliferation and the mechanisms involved. All 3 members of the K(Ca) channel family were found in fibroblasts. 1,3-Dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2H-benzimidazol-2-one (NS1619, a BK channel activator) or 1-ethyl-2-benzimidazolinone (EBIO, an IK channel activator) decreased the proliferation of fibroblasts and induced apoptotic changes by mitochondrial membrane potential disruption. However, a pan-caspase inhibitor (Z-VAD-fmk) failed to prevent the apoptotic changes. Our findings indicate that 3 types of functional K(Ca) channels are expressed in human dermal fibroblasts and are involved in apoptosis of the cells through the mitochondrial apoptotic pathway, but seemingly in a caspase-independent manner.


[The use of biocompatible materials in apicoectomy].
The use of more biocompatible materials in comparison with silver, gutta-percha and amalgam was introduced a decade ago. First titanium was used, later followed by alumina. Both clinical as well as radiological results favor their use. Yet it appears that the system is mainly confined to the maxillary front teeth and mandibular canines.


Sutureless Technique to Fix the Great Saphenous Vein along the Atrioventricular Groove Using Fibrin Glue in Off-Pump Coronary Artery Bypass Grafting.
We describe a simple method to fix the great saphenous vein graft (SVG) to the right coronary artery along the atrioventricular groove using fibrin glue in off-pump coronary artery bypass grafting (OPCAB). After completion of the proximal anastomosis, the SVG was placed along the atrioventricular groove to the acute margin. Fibrin glue was sprayed using pressurized carbon dioxide gas. A distal anastomosis was subsequently performed after rotating the heart to expose the posterior descending artery. It is a straightforward and reproducible technique to determine the optimal length of the SVG and prevent kinking or stretching of the graft, especially in OPCAB.


Human MATER localization in specific cell domains of oocytes and follicular cells.
MATER (Maternal Antigen That Embryos Require) is an oocyte-specific protein dependent on the maternal genome and required for early embryonic development. The gene products expressed in oocytes play important roles in folliculogenesis, fertilization and pre-implantation development. The aim of this study was to characterize the localization and distribution pattern of the human MATER protein during follicular development and after ovulation, to determine its functional role. Immunocytochemistry experiments coupled with confocal and electron microscopy analysis were carried out to determine the ultrastructural localization of MATER in human ovarian tissue and in isolated oocytes, obtained during IVF procedures. Human cumulus cells were cultured, with or without cycloheximide, to confirm endogenous biosynthesis of the protein. Human MATER is detectable at the onset of the follicular maturation process, suggesting this protein has a role at earlier stages in the human compared with other mammalian species. The presence of MATER is specific to the oocyte and follicular cells that, during maturation, are spatially and functionally associated with the oocyte. The nuclear, nucleolar and mitochondrial localization hints at a possible role in RNA processing and the metabolic activity of the cell.


A potential role for multiple tissue kallikrein serine proteases in epidermal desquamation.
Desquamation of the stratum corneum is a serine protease-dependent process. Two members of the human tissue kallikrein (KLK) family of (chymo)tryptic-like serine proteases, KLK5 and KLK7, are implicated in desquamation by digestion of (corneo)desmosomes and inhibition by desquamation-related serine protease inhibitors (SPIs). However, the epidermal localization and specificity of additional KLKs also supports a role for these enzymes in desquamation. This study aims to delineate the probable contribution of KLK1, KLK5, KLK6, KLK13, and KLK14 to desquamation by examining their interactions, in vitro, with: 1) colocalized SPI, lympho-epithelial Kazal-type-related inhibitor (LEKTI, four recombinant fragments containing inhibitory domains 1-6 (rLEKTI(1-6)), domains 6-8 and partial domain 9 (rLEKTI(6-9')), domains 9-12 (rLEKTI(9-12)), and domains 12-15 (rLEKTI(12-15)), secretory leukocyte protease inhibitor, and elafin and 2) their ability to digest the (corneo)desmosomal cadherin, desmoglein 1. KLK1 was not inhibited by any SPI tested. KLK5, KLK6, KLK13, and KLK14 were potently inhibited by rLEKTI(1-6), rLEKTI(6-9'), and rLEKTI(9-12) with Ki values in the range of 2.3-28.4 nm, 6.1-221 nm, and 2.7-416 nm for each respective fragment. Only KLK5 was inhibited by rLEKTI(12-15) (Ki = 21.8 nm). No KLK was inhibited by secretory leukocyte protease inhibitor or elafin. Apart from KLK13, all KLKs digested the ectodomain of desmoglein 1 within cadherin repeats, Ca2+ binding sites, or in the juxtamembrane region. Our study indicates that multiple KLKs may participate in desquamation through cleavage of desmoglein 1 and regulation by LEKTI. These findings may have clinical implications for the treatment of skin disorders in which KLK activity is elevated.


Combination of the (87)Sr/(86)Sr ratio and light stable isotopic values (δ(13)C, δ(15)N and δD) for identifying the geographical origin of winter wheat in China.
This study aims to investigate whether isotopic signatures can be used to develop reliable fingerprints for discriminating the geographical origin of Chinese winter wheat, and to evaluate the discrimination effects of δ(13)C, δ(15)N and δD, alone or with (87)Sr/(86)Sr. In this study, the values of δ(13)C, δ(15)N and δD, and the (87)Sr/(86)Sr ratios of wheat and provenance soils from three regions were determined. Significant differences were found in all parameters of wheat and (87)Sr/(86)Sr in soil extract (reflecting the bioavailable fraction of soil) among different regions. A significantly positive correlation was observed between the (87)Sr/(86)Sr ratios of wheat and soil extracts. An overall correct classification rate of 77.8% was obtained for discriminating wheat from three regions based on light stable isotopes (δ(13)C, δ(15)N, and δD). The correct classification rate of 98.1% could be obtained with the combination of the (87)Sr/(86)Sr ratio and the light stable isotopic values.


Serum estradiol and risk of stroke in elderly men.
To determine if levels of serum estradiol and testosterone can predict stroke in a population-based sample of elderly men. Serum 17beta estradiol and testosterone were measured in 2,197 men aged 71 to 93 years who participated in the Honolulu-Asia Aging Study from 1991 to 1993. All were free of prevalent stroke, coronary heart disease, and cancer. Participants were followed to the end of 1998 for thromboembolic and hemorrhagic events. During the course of follow-up, 124 men developed a stroke (9.1/1,000 person-years). After age adjustment, men in the top quintile of serum estradiol (> or =125 pmol/L [34.1 pg/mL]) experienced a twofold excess risk of stroke vs men whose estradiol levels were lower (14.8 vs 7.3/1,000 person-years, p < 0.001). Among the lower quintiles, there were little differences in the risk of stroke. Findings were also significant and comparable for bioavailable estradiol and for thromboembolic and hemorrhagic events. After additional adjustment for hypertension, diabetes, adiposity, cholesterol concentrations, atrial fibrillation, and other characteristics, men in the top quintile of serum estradiol continued to have a higher risk of stroke vs those whose estradiol levels were lower (relative hazards = 2.2; 95% CI = 1.5 to 3.4, p < 0.001). Testosterone was not related to the risk of stroke. High levels of serum estradiol may be associated with an elevated risk of stroke in elderly men.


Cell membrane stretch modulates the high-conductance Ca2+-activated K+ channel in bovine trabecular meshwork cells.
Anterior chamber structures are subjected to changes in intraocular pressure (IOP). Several studies have pointed out that trabecular meshwork (TM) cells are sensitive to mechanical stretch and that cell-signaling mechanisms are activated in response to elevated pressure. Because membrane stretch has been shown to be a modulator of several ionic conductances, this study was conducted to determine its effects on the high-conductance Ca(2+)-activated K(+) (BK(Ca)) channels present in TM cells. Primary cultures of TM cells from bovine eyes were used. Patch-clamp recordings were performed in the cell-attached, inside-out, and whole-cell configurations. To stretch the cell membrane, both suction to the rear end of the patch pipette and hypotonic shock were used. Intracellular calcium concentration ([Ca(2+)](i)) was measured in TM cells loaded with fura-2, using an epifluorescence microscope coupled to a charge-coupled device (CCD) camera. Electrophysiological characterization of BK(Ca) channels was in agreement with previous studies. In cell-attached patches, the open probability of the BK(Ca) channel (i.e., the amount of time the channel is open) increased consistently when 14- to 45-mm Hg suctions were applied at a constant depolarized voltage. At a constant pressure (25 or 45 mm Hg), channel openings increased when depolarizing pulses were applied to the patch. Stretch activation of the BK(Ca) channel was not mediated by increases in [Ca(2+)](i), because it was present in inside-out patches maintained at a constant Ca(2+) concentration. Nevertheless, it cannot be ruled out that at low suction levels, a minimum Ca(2+) concentration is necessary for channel activation. Whole-cell currents carried by BK(Ca) channels increased when the isotonic solution in the bath was exchanged with a hypotonic solution and were selectively blocked by iberiotoxin. In our conditions, the hypotonic shock did not modify [Ca(2+)](i). The data show that in TM cells, open probability of the BK(Ca) channel is enhanced by membrane stretching as well as by membrane depolarization and [Ca(2+)](i). Changes in membrane tension induced by cell volume increase also activated whole-cell BK(Ca) currents. Homeostatic mechanisms in TM cells may involve BK(Ca) channel activation in response either to changes in cell volume or changes in IOP.


Decreased level of interleukin-10-producing B cells in patients with pemphigus but not in patients with pemphigoid.
While the frequency of interleukin (IL)-10-producing B (B10) cells is reported to have an inverse correlation with disease activity in some human autoimmune diseases, the association between B10 cells and autoimmune blistering diseases (AIBD) has not been well evaluated. Although several phenotypes of human regulatory B cells have been proposed, the most appropriate one in AIBD has not been established. To evaluate B10 cells in AIBD including their phenotypes. Peripheral blood mononuclear cells were isolated from 39 patients with AIBD, including 14 with pemphigus and 25 with pemphigoid, and 10 healthy controls. We investigated the frequencies of B10 cells and CD19+ CD24hi CD38hi B cells using flow cytometry. The frequencies of B10 cells and CD19+ CD24hi CD38hi B cells were significantly lower and higher, respectively, in patients with pemphigus compared with healthy controls. Comparing patients with pemphigoid and healthy controls, no significant difference in the frequencies of B10 cells and CD19+ CD24hi CD38hi B cells was observed. B10-cell level in pemphigus was not associated with disease severity but inversely correlated with the required dose of steroid for treatment. While no significant difference in the frequency of IL-10-producing cells among CD19+ CD24hi CD38hi B cells was observed, in CD9+ and CD27- B-cell subsets it was significantly decreased in patients with pemphigus compared with healthy controls. Our results suggest the association of B10 cells with pemphigus but not with pemphigoid. The decrease in B10-cell level in pemphigus is partly caused by the lower production of IL-10 in CD9+ and CD27- B-cell subsets.


Posttraumatic stress disorder among adolescent earthquake victims in Taiwan.
To assess the exposure experience and prevalence of posttraumatic stress disorder (PTSD) among adolescent victims in the worst-affected region (Chungliao) near the epicenter of a severe earthquake (7.3 on the Richter scale) that occurred on September 21, 1999, in Taiwan. The experience of exposure to the earthquake and subjective symptoms of junior high school students aged 12 to 14 who remained in the area were assessed with self-rated questionnaires. Psychiatrists made independent diagnoses for PTSD by using the Children's Interview for Psychiatric Syndromes. Six weeks after the earthquake, 21.7% of 323 students demonstrated PTSD. Those with PTSD showed significantly more psychiatric symptoms than did those without PTSD. Being physically injured and experiencing the death of a close family member with whom they had lived were the 2 major risk factors for PTSD. This study demonstrates that PTSD among adolescent victims of a severe earthquake in Taiwan is not as high as that reported in other studies. Methodological differences in the investigations are discussed, along with differences in symptom manifestations. However, long-term follow-up of these victims is recommended to prevent the development of other psychiatric complications.


Effect of carbohydrates and night temperature on night respiration in rice.
Global warming causes night temperature (NT) to increase faster than day temperature in the tropics. According to crop growth models, respiration incurs a loss of 40-60% of photosynthate. The thermal sensitivity of night respiration (R(n)) will thus reduce biomass. Instantaneous and acclimated effects of NT on R(n) of leaves and seedlings of two rice cultivars having a variable level of carbohydrates, induced by exposure to different light intensity on the previous day, were investigated. Experiments were conducted in a greenhouse and growth chambers, with R(n) measured on the youngest fully expanded leaves or whole seedlings. Dry weight-based R(n) was 2.6-fold greater for seedlings than for leaves. Leaf R(n) was linearly related to starch (positive intercept) and soluble sugar concentration (zero intercept). Increased NT caused higher R(n) at a given carbohydrate concentration. The change of R(n) at NT increasing from 21 °C to 31 °C was 2.4-fold for the instantaneous response but 1.2- to 1.7-fold after acclimation. The maintenance component of R(n) (R(m)'), estimated by assimilate starvation, averaged 28% in seedlings and 34% in leaves, with no significant thermal effect on this ratio. The acclimated effect of increased NT on R(m)' across experiments was 1.5-fold for a 10 °C increase in NT. No cultivar differences were observed in R(n) or R(m)' responses. The results suggest that the commonly used Q10=2 rule overestimates thermal response of respiration, and R(n) largely depends on assimilate resources.


Density functional theory for sphere-needle mixtures: toward finite rod thickness.
For mixtures of hard spheres and hard spherocylinders of large aspect ratio a recently proposed density functional theory is extended to incorporate effects due to nonvanishing rod thickness. This is accomplished by introducing several new geometric weight functions into the framework. We demonstrate explicitly how these weight functions recover the sphere-rod Mayer bond.


Abdominoplasty following colostomy.
An abdominoplasty was performed on a 57-year-old woman, 5 years after she had undergone a permanent colostomy for ulcerative colitis. The abdominoplasty was performed in an attempt to reduce the increased abdominal girth thought to be responsible for excess leakage of effluent from the colostomy site with changes in posture and increased abdominal pressure. A simple abdominoplasty flap was designed to decrease the excess abdominal tissue without impinging on the colostomy site. The result was dramatic in accomplishing the set-out goals. An added bonus was a reversal of preoperative urinary incontinence. This outcome is thought to be a result of more efficient voiding after abdominal muscle plication and stabilization.


The regulation of vestibular afferent information during monocular vision while standing.
The purpose of this study was to investigate the contribution of the vestibular system to postural control during monocular vision using binaural-bipolar galvanic vestibular stimulation (GVS). Four visual (both eyes, dominant eye, non-dominant eye, and no vision) conditions were tested during GVS in five healthy subjects while focusing on a target placed in front of them. GVS evoked similar upper body postural sway during both monocular and no vision conditions that were significantly greater to those during binocular vision. Changes in ground reaction forces to the anode side followed that same trend, although data for vision with the dominant eye were not significantly different from that for binocular vision. These data suggest an increase in the weighting of vestibular afferent information during monocular vision for standing postural control.


Adenocarcinoma of a colostomy following abdominoperineal resection for squamous cell carcinoma of the anal canal: a case study.
Malignant neoplasms presenting on a stoma, as well as the development of colorectal adenocarcinoma after previous treatment for squamous cell carcinoma (SCC) of the anal canal, are rare. The unique case is presented of an 81-year-old woman with parastomal bleeding and ulceration found to have a primary colorectal adenocarcinoma arising de novo on a colostomy, formed after salvage abdominoperineal resection (APR) 3 years earlier for recurrent anal SCC. This is the first reported case of a colonic adenocarcinoma on a colostomy formed after an APR for anal SCC. Although stomal neoplasia is rare, the appearance of a friable bleeding lesion on the stoma should be investigated to exclude metastatic cancer or a second primary malignancy.


Maternal dietary intake in twin pregnancies: does it diminish towards term?
We tested the hypothesis that energy intake among women with twin pregnancies decreases in late gestation, as the uterus enlarges and they become increasingly uncomfortable. We asked women to recall dietary intake for the previous 24 hours, every 2 weeks from around 29 weeks of gestation, using a photographic atlas and other strategies to estimate portion size. Eighty women provided data at around 29, 31, 33 and 35 weeks of gestation. We calculated total energy intake, and energy intake from carbohydrate, fat and protein. Data were log transformed to reduce skewness. Using mixed effects linear regression models, we found minimal evidence that total energy intake changed over this time period, either before or after adjustment for potential confounding factors (95% confidence interval for relative change per week: -1.4%, 0.6%). There was weak evidence of a small decline in carbohydrate intake over time, both before and after controlling for energy intake. We cannot exclude change in energy intake or diet composition before 29 weeks or after 35 weeks of twin gestation.


Protein synthesis and the mechanisms of lasting change in anxiety induced by severe stress.
Brief, unprotected exposure of rats to cats (predator stress) may be lastingly anxiogenic in a variety of tests of rodent anxiety. Recent findings suggest that predator stress induced plasticity in neural circuitry implicated in fear learning underlies some of these anxiogenic effects. In addition, recent work implicates a consolidation-like process in the impact of predator stress on anxiety in that effects of predator stress may be interrupted by immediate post stressor pharmacological interventions. The present study tested whether "consolidation" of the anxiogenic effects of predator stress were dependent on protein synthesis. In addition, the study examined whether a protein synthesis dependent reconsolidation-like process was at work when rats were exposed to a cat twice. Anisomycin (210 mg/kg) or vehicle (Tween 80 in saline) was injected subcutaneously 1 min after a single cat exposure (consolidation test paradigm) or a 1 min after a second cat exposure (reconsolidation test paradigm) and behavior tested 7-8 days after predator stress. In the consolidation test paradigm, anisomycin blocked the anxiogenic effects of predator stress in the elevated plus maze (EPM) measured with open arm exploration. Moreover, anisomycin blocked the potentiation of startle by predator stress when rats were startled in the light, but not when startled in the dark. In contrast, the delay of habituation of startle produced by predator stress was unaffected by anisomycin. Suppression of risk assessment in the EPM by predator stress was not affected by anisomycin either. In startle testing, vehicle injection 1 min after predator stress led to a lasting suppression, rather than enhancement of startle response. Vehicle plus predator stress enhanced and prolonged corticosterone level changes sampled over 30-180 min after treatment when compared to handled or predator stressed only rats. In addition, predator stress plus vehicle suppression of startle was blocked by a benzodiazepine anxiolytic (chloradiazepoxide) or the glucorticoid receptor (GR) blocker RU486. Both drugs returned startle to the predator stressed only heightened levels. It is argued that an added anxiogenic effect of vehicle injection plus predator stress leads to a suppression, rather than enhancement of startle. Startle suppression appears to be mediated, in part, by activation of GR by corticosterone which engages a protein synthesis dependent process, since anisomycin blocked the startle suppressive effects of vehicle. Startle suppression also appeared to be independent of the startle enhancing effect of predator stress and in competition with it. Since predator stress may model aspects of hyperarousal associated with post traumatic stress disorder (PTSD), implications of these findings for understanding of mechanisms of initiation of the disorder and for treatment are discussed.


Trends in contraceptive use among Florida women: implications for policies and programs.
Our study objective was to assess changes in effective contraceptive use among women at risk of unintended pregnancy in Florida in 2008 and 2009 compared with 2002 and 2004. Contraceptive use questions were available from Florida's Behavioral Risk Factor Surveillance System (BRFSS) for both periods (n = 4,606). Log binomial regression was used with appropriate methods to account for complex sampling in the BRFSS. We examined the change in four effective contraceptive use groups: sterilization, long-acting reversible contraceptive (LARC), short-acting reversible contraceptive (SARC), and barrier methods. Prevalence ratios comparing the two time periods were adjusted by demographic characteristics, employment, insurance status, children at home, poverty level, health behaviors, and health status. No evidence of change was found in sterilization (Adjusted Prevalence Ratio APR = 0.96; 95 % CI: 0.84-1.10) or SARC (APR = 1.01; 95 % CI: 0.87-1.18). The overall use of LARC increased and use of barrier methods decreased significantly over the two periods (APR = 1.68; 95 % CI: 1.09-2.60 and APR = 0.77; 95 % CI: 0.61-0.98, respectively). Only two population groups experienced significant changes in prevalence in the four use groups over this period. Non-Hispanic White women increased their use of LARC (APR = 2.89; 95 % CI: 1.58-5.29) and women who have never been married decreased their use of barrier methods (APR = 0.51; 95 % CI: 0.33-0.77). Contraceptive use in Florida continues to be low overall with some shift towards more effective long-term methods. New efforts are needed to promote and increase family planning practices, which include the use of effective contraceptives.


Panclavicular ankylosis in pustulotic arthroosteitis. A case report.
A 54-year-old man who had palmoplantar pustulosis showed bilateral, complete osseous fusion of the sternoclavicular and acromioclavicular joints. No significant abnormality was seen in the glenohumeral joint. The left clavicle had a nonunion develop, whereas the right did not. The elevation of the right shoulder was limited to 100 degrees in active and passive measurements. The total external rotation and internal rotation at the side was 135 degrees. This decreased to 90 degrees at 60 degrees elevation and to 10 degrees at maximum (100 degrees) elevation. Based on the kinematic data on normal shoulders, it was thought that the ankylosis of both ends of the clavicle held the scapula unrotated during the arm movement so that, at the arm to trunk angle of 100 degrees, the position of the humerus relative to the scapula was equivalent to that of a normal shoulder in complete elevation. The current case provided an extremely rare clinical setting where shoulder mobility depended only on glenohumeral motion as a result of the complete loss of scapulothoracic motion.


Wnt/beta-catenin signaling mediates oval cell response in rodents.
Adult hepatic stem cells or oval cells are facultative stem cells in the liver that are activated during regeneration only during inhibition of innate hepatocyte proliferation. On the basis of its involvement in liver cancer, regeneration, and development, we investigated the role of the Wnt/beta-catenin pathway in oval cell response, which was initiated in male Fisher rats with 2-acetylaminofluorine and two-third partial hepatectomy (PHX). Extensive oval cell activation and proliferation were observed at 5 and 10 days post-PHX, as indicated by hematoxylin-eosin and proliferating cell nuclear antigen analysis. A noteworthy increase in total and active beta-catenin was observed at this time, which was localized to the oval cell cytoplasm and nuclei by immunohistochemistry and confirmed by double immunofluorescence. A concomitant increase in Wnt-1 in hepatocytes along with increased expression of Frizzled-2 in oval cells was observed. This paracrine mechanism coincided with a decrease in Wnt inhibitory factor-1 and glycogen synthase kinase-3beta down-regulation leading to beta-catenin stabilization. To strengthen its role, beta-catenin conditional knockout mice were treated with 3,5-diethoxycarbonyl-1,4-dihydrocollidine to induce oval cell activation. A dramatic decrease in the A6-positive oval cell numbers in the absence of beta-catenin demonstrated a critical role of beta-catenin in oval cell biology. The Wnt/beta-catenin pathway plays a key role in the normal activation and proliferation of adult hepatic stem cells.


Caroli's syndrome and adult polycystic kidney disease.
Caroli's disease is a cystic disease of the liver, which has been rarely associated with adult onset polycystic kidney disease. Three cases have been reported in the English Medline search. The presentation of this fourth case discusses the issues surrounding the treatment of Caroli's disease in the setting of a renal transplant.


16-electron elongated dihydrogen complex stabilized by agostic interaction.
A 16-electron dicationic dihydrogen complex [Ru(eta2-H...H)(PP)2][OTf]2 [4; PP = (C6H5CH2)2PCH2CH2P(CH2C6H5)2] has been prepared and characterized by protonating the precursor hydride complex [Ru(H)(PP)2)][OTf] (2) using HOTf. The hydride and dihydrogen complexes are stabilized via agostic interaction of the ortho C-H fragment of the phenyl ring on the benzyl group. The intact nature of the H-H bond in this derivative was established from the short spin-lattice relaxation time and the observation of a substantial J(H,D) of 22.0 Hz for the HD isotopomer. The H-H bond distance calculated from J(H,D) is 1.05 A, which falls under the category of elongated dihydrogen ligands.


Comparison of two mononuclear cell program settings on two apheresis devices intended to collect high yields of CD14+ and CD3+ cells.
In cancer and transplantation therapy apheresis devices and software of optimum standards are required for the collection of high cell yields with high purity of the desired cell fraction. In a paired study, 15 healthy blood donors underwent four 10-L leukapheresis procedures (197 +/- 33 min) with an inlet blood flow rate of 60 mL per minute by use of two different MNC program settings of the COBE Spectra (Gambro BCT) and the AS.TEC 204 (Fresenius Hemocare) cell separators. Use of the standard MNC program of both apheresis devices resulted in significantly higher (p < 0.01) collection efficiencies of CD14+ monocytes, CD3+ cells, CD4+ cells, CD8+ T cells, CD16+ CD56+ natural killer (NK) cells, and residual PLTs (p < 0.001), owing to higher centrifuge speed. The mean MNC purity of all components was more than 90 percent. By use of standard programs of either device, significant correlations (p < 0.01) between donor monocytes and preleukapheresis NK cell counts and the corresponding component cell yields were found. Compared to the program modifications with lower centrifuge velocities the standard MNC programs were significantly more efficient regarding CD14+, CD3+, and CD16+ CD56+ cells. Enhanced centrifuge speed and inlet blood flow rate in MNC programs resulted in higher, similar composed MNC concentrations of the products.


The significance of suicide notes.
Studying the contents of suicide notes might help to improve the understanding of individual cases whilst offering an opportunity to search for possible preventive guidelines. In a study in an Australian capital city, suicides for one year (1972-73) were investigated including the medical and psychiatric data where possible, and psychosocial circumstances in each case. The total number of suicides in the year was 135. Twenty-seven individuals left suicide notes of which 25 were available for examination. The mood as reflected in the notes was categorised in terms of depression and hopelessness, neutral affect and hostility. The affect was related to age, sex, marital status and method of suicide. The results are presented with examples of each category and are discussed with respect to their contribution to (i) understanding the cause of death, (ii) understanding the intention of the suicide, (iii) assisting the relatives in coping with the suicide, (iv) drawing attention to those factors where earlier intervention may have averted suicide.


Coagulation, inflammation and myocardial dysfunction in unstable coronary artery disease and the influence of glycoprotein IIb/IIIa inhibition and low molecular weight heparin.
Patients with unstable coronary artery disease (CAD) have an increased risk of subsequent myocardial infarction and death. This study evaluated the safety and efficacy of treatment with glycoprotein IIb/IIIa inhibition in addition to aspirin, low molecular-weight heparin and its influence on coagulation and inflammation. Also, early and differentiated risk assessment utilising markers of inflammation, myocardial damage and dysfunction were evaluated. The Global Utilisation of Strategies To open Occluded arteries-IV (GUSTO-IV) trial randomised 7800 patients with unstable CAD to 24 or 48 hours infusion of abciximab or placebo in addition to routine treatment with aspirin and heparin or dalteparin. Baseline levels of creatinine, C-reactive protein (CRP), troponin T (TnT) and N-terminal pro-brain natriuretic peptide (NT-proBNP) were analysed. At selected sites, all patients received subcutaneous dalteparin (n=974), in stead of heparin infusion (n=6826). In a sub-population of dalteparin treated patients (n=404), serial measurements of markers of coagulation, fibrinolysis and inflammation were also performed. Addition of abciximab to dalteparin as the primary treatment of unstable CAD was not associated with any significant reduction in cardiac events but a doubled risk of bleedings. The combination of abciximab with dalteparin seemed as safe when used with heparin. Despite full dose dalteparin and aspirin there was a simultaneous activation of the inflammation, coagulation and fibrinolysis systems without any influence of the abciximab treatment. Elevated levels of CRP, TnT, and NT-proBNP and reduced creatinine clearance were independently related to short and long-term mortality. The best prediction of high and low risk was provided by a combination of NT-proBNP and creatinine clearance. Any detectable elevation of TnT and reduced creatinine clearance, but neither elevation of CRP nor NT-proBNP, were also independently associated to a raised risk of subsequent myocardial infarction.


Surgical hysteroscopic treatment of cesarean-induced isthmocele in restoring fertility: prospective study.
The reproductive outcome in 41 consecutive patients with cesarean-induced isthmocele and secondary infertility was evaluated prospectively. Patients included menopausal women (mean [SD; 95% CI] age, 35 [4.1; 29-42] years), with fertility duration of 3 to 8 (4.6 [28]) years with isthmocele, postmenstrual abnormal uterine bleeding, and suprapubic pelvic pain. Transvaginal ultrasound and office hysteroscopy were used to diagnosis isthmocele. Complete fertility tests were performed to exclude other causes of infertility in both female and male participants. Operative hysteroscopy was performed to correct the cesarean scar defect, and histologic findings were evaluated. Correction of isthmocele via operative hysteroscopy was successful in all cases evaluated. Patients became pregnant spontaneously between 12 and 24 months after isthmoplasty. Thirty-seven of the 41 patients (90.2%) delivered via cesarean section, and 4 (9.8%) had a spontaneous abortion in the first trimester. Isthmoplasty resulted in resolution of postmenstrual abnormal uterine bleeding and suprapubic pelvic pain in all patients. Thus, it was concluded that surgical treatment of cesarean-induced isthmocele using a minimally-invasive approach (operative hysteroscopy) restores fertility and resolves symptoms in women with a cesarean section scar and secondary infertility.


Work-related Helicobacter pylori infection among sewage workers in municipal wastewater treatment plants in Belgium.
Most studies among sewage workers of wastewater treatment plants (WWTPs) have found a higher prevalence of gastrointestinal symptoms than among non-sewage exposed workers. Waterborne transmission of Helicobacter pylori (H pylori) has been hypothesised, as the bacteria can survive into an aqueous environment and has been detected in sewage. A health and hygiene questionnaire has demonstrated a higher prevalence of peptic ulcers among Belgian WWTP operators and maintenance workers than among non-sewage exposed colleagues. To assess the seroprevalence of H pylori infection in Belgian sewage workers at municipal WWTPs, and to determine whether sewage exposure is an important risk factor for acquisition of H pylori and the possible association with gastrointestinal symptoms. A seroprevalence study of H pylori antibodies was conducted among 317 WWTP employees (operators, maintenance workers, laboratory personnel, other job). Information about demographic variables, possible H pylori risk factors, working history, and history of current gastrointestinal symptoms during last 3 months was obtained by a questionnaire. The presence of H pylori IgG was investigated with an ELISA. The results were compared with those of 250 employees of a pharmaceutical company (operators, maintenance workers, laboratory personnel). The prevalence of H pylori IgG antibodies among sewage workers was 16.7% (95% CI 12.6 to 20.8%) compared to 13.6% (95% CI 9.4 to 17.8%) among the control group. In a logistic regression model with controlling for age and educational levels, OR study/control group was 1.02 (95% CI 0.58 to 1.80 with p=0.93). No significant associations were found between the H pylori status and gastrointestinal symptoms, occupational exposures in different tasks, nor with hygienic practices. Our results do not suggest that H pylori infection is a probable cause of part of gastrointestinal symptoms among Flemish municipal sewage workers of WWTPs.


Effects of methylprednisolone on peritumoral brain edema. A quantitative autoradiographic study.
Peritumoral brain edema was produced by intracerebral transplantation of Walker 256 tumor in rats. Local cerebral blood flow (LCBF), local cerebral glucose utilization (LCGU), and capillary permeability were studied in untreated and methylprednisolone-treated rats by quantitative autoradiography. In the untreated group, LCBF and LCGU were widely depressed in the cortex and deep structures of the hemisphere ipsilateral to the tumor. In the methylprednisolone-treated animals, LCBF and LCGU were significantly better than in the untreated animals. Capillary permeability was highly increased in the viable part of the tumor in the untreated animals. In the methylprednisolone-treated group, capillary permeability of the tumor was significantly lower than that in the untreated group. These results may suggest that increase in capillary permeability of the tumor is the major source for edema fluid production, and that methylprednisolone improves brain edema by decreasing capillary permeability of the tumor. Decrease in edema fluid formation may result in restoration of blood flow and glucose metabolism in the adjacent brain tissue, and may improve clinical symptoms and signs.


Neuroprotection in focal cerebral ischemia owing to delayed treatment with melanocortins.
In gerbils subjected to transient global cerebral ischemia, melanocortin peptides produce long-lasting protection with a broad time window, and through the activation of central nervous system melanocortin MC(4) receptors. Here we aimed to investigate whether melanocortins are neuroprotective also in a rat model of focal cerebral ischemia induced by intrastriatal microinjection of endothelin-1. The vasoconstrictor agent endothelin-1 caused a significant impairment in spatial learning and memory, as well as in sensory-motor orientation and limb use, associated with severe striatal morphological damage including intense neuronal death and an almost complete myelin degradation. Treatment of ischemic rats with a nanomolar dose (340 microg/kg/day i.p. for 11 days, beginning 3 h or 9 h after endothelin-1 microinjection) of the melanocortin analog [Nle(4), D-Phe(7)]alpha-melanocyte-stimulating hormone (NDP-alpha-MSH) significantly reduced striatal damage, and improved subsequent functional recovery, with all scheduled NDP-alpha-MSH treatments. Pharmacological blockade of melanocortin MC(4) receptors prevented the protective effect of NDP-alpha-MSH. Our findings give evidence that melanocortins are neuroprotective, with a broad time window, also in a severe model of focal cerebral ischemia, and suggest that melanocortin MC(4) receptor agonists could produce neuroprotection in different experimental models of ischemic stroke.


Does the efficacy of parent-child training depend on maternal symptom improvement? Results from a randomized controlled trial on children and mothers both affected by attention-deficit/hyperactivity disorder (ADHD).
Multimodal treatment of children with ADHD often includes parent-child training (PCT). However, due to the high heritability, parents of children with ADHD are frequently also affected by the disorder, which is likely to constitute a significant barrier to successful treatment of the child. This secondary analysis of our randomized controlled multicentre AIMAC trial (ADHD in mothers and children) investigates whether children's outcomes following parent-child training in combination with maternal ADHD treatment depend on maternal symptom improvement. In a first step focusing on treatment of maternal ADHD, 144 mothers of mother-child dyads were randomized to multimodal ADHD treatment (group psychotherapy plus methylphenidate) or clinical management (mainly supportive counselling). After 12 weeks (T2), a 12-week PCT program (T2-T3) for all mother-child dyads was added to treat children's ADHD. Maternal symptomatology (CAARS-O:L; SCL-90-R) and children's externalizing symptoms (ADHD-ODD Scale, SDQ) were repeatedly assessed (T1 = baseline, T2, T3). Effects of changes in maternal symptomatology (T1-T2) on the change in children's symptom scores (T1-T3) were analysed using a general linear model, controlling for baseline scores, study centre, and maternal treatment group. 125 mother-child dyads were analysed. Mothers showed significant improvements in ADHD symptoms and overall psychopathology [CAARS-O:L ADHD index: mean - 3.54, SE 0.74 p < 0.0001; SCL-90-R Global Severity (GS): mean - 11.03, SE 3.90, p = 0.0056]. Although children's externalizing symptoms improved significantly (ADHD-ODD Scale: mean - 4.46, SE 0.58, p < 0.0001), maternal improvement had no effect on children's outcomes after Bonferroni-Holm correction for multiple testing. The findings do not support our hypothesis that children's outcomes following PCT for ADHD depend on maternal symptom improvements.Trial register CCT-ISRCTN73911400.


Deuterium dilution technique for body composition assessment: resolving methodological issues in children with moderate acute malnutrition.
Childhood malnutrition is highly prevalent and associated with high mortality risk. In observational and interventional studies among malnourished children, body composition is increasingly recognised as a key outcome. The deuterium dilution technique has generated high-quality data on body composition in studies of infants and young children in several settings, but its feasibility and accuracy in children suffering from moderate acute malnutrition requires further study. Prior to a large nutritional intervention trial among children with moderate acute malnutrition, we conducted pilot work to develop and adapt the deuterium dilution technique. We refined procedures for administration of isotope doses and collection of saliva. Furthermore, we established that equilibration time in local context is 3 h. These findings and the resulting standard operating procedures are important to improve data quality when using the deuterium dilution technique in malnutrition studies in field conditions, and may encourage a wider use of isotope techniques.


Aminoglycosides: another perspective.
Despite the introduction of several new classes of antimicrobial agents, aminoglycosides are still recognized as first-line therapeutic agents in the management of severe gram-negative sepsis. The major obstacle limiting the use of aminoglycoside antibiotics has been, and continues to be, the possibility of drug-induced ototoxicity and nephrotoxicity. This review critically examines the definitions used to establish the diagnosis of aminoglycoside-induced nephrotoxicity and ototoxicity and the clinical significance of these adverse reactions. The review also focuses on the practical and economic issues surrounding therapeutic drug monitoring practices. We conclude that aminoglycoside antibiotics remain an effective and economical form of therapy for severe infections and that if careful criteria are used in the selection of these agents, the benefits of therapy outweigh the risk of toxicity.


[Important vaccine accidents in history of vaccine and their apocalypses].
Although it has been widely acknowledged that glorious history of vaccination of past 200 years has been a successful feat in the protection of multiple disease calamities, contamination, incomplete attenuation or high virulence of the vaccine, laboratory regulation, injection and preservation problems have witnessed many accidents in the river of history. This review would summarize and comment on typical events of accidents regarding vaccines, within the framework designed to provide experience and apocalypses


[Stereotactic surgery in Parkinson disease: patient selection criteria in the light of existing research].
The renewal of interest in the neurosurgical treatment of Parkinson's disease (PD) also in Poland results from of our improved understanding of functional anatomy of basal ganglia, developments in neurophysiological and neuroimaging techniques, as well as from advances in stereotactic surgery techniques. In view of the growing number of PD patients referred to surgical treatment and the wide variety of interventions offered, the development of clear of PD patient's selection to surgery seems necessary. Various surgical options and possible targets provide different functional benefits, but the almost 10 year's experience makes us aware also of the limitations and possible complications involved. Algorithms worked out by researchers from the most experienced centers include the following selection criteria as a minimal standard of the PD patient's evaluation before surgery: a reliable diagnosis of Parkinson's disease, at least 5 years of PD duration since the onset of symptoms, good responsiveness to L-dopa or apomorphine, exclusion of severe depression and dementia, neuroimaging (MRI) performed before the surgery, and optimal (but ineffective) attempts at available pharmacological therapy prior to the surgery.


Lymph node metastasis, recurrence, and prognosis in small peripheral lung adenocarcinoma. Analysis based on replacement.
We studied clinical and pathological features of small peripheral adenocarcinoma of the lung, focusing on tumor typing based on Noguchi's classification of small adenocarcinoma and determining whether these tumors grew by replacing alveolar lining cells. Subjects were 51 patients with small peripheral adenocarcinoma 2 cm or less in diameter resected between 1994 and 2001. Mediastinal and hilar lymph node dissection was done in 37 (72.5%). Patients were divided into 2 groups by replacement or nonreplacement tumors. We compared patient profiles, lymph node involvement, and recurrence and survival patterns. No significant difference was seen between groups in mean age, surgical procedure, or primary tumor location. Women predominated in replacement tumors at 71% vs 41%, p = 0.04. The incidence of lymph node metastasis at 40% vs 4.5%, p = 0.007 and distant metastasis at 47% vs 2.9%, p < 0.001 was significantly higher in nonreplacement than replacement tumors. Replacements tumor thus showed significantly better disease-free survival at 95% vs 53%, p < 0.001, and overall 3-year survival at 95.4% vs 62.7% than did nonreplacement tumors. We found distant metastasis and lymph node involvement to be more frequent in nonreplacement than replacement small peripheral adenocarcinoma, on suggesting that pretreatment tumor typing and accurate nodal status determination are essential to improve disease staging.


Isolation of recombinant viruses between cauliflower mosaic virus and a viral gene in transgenic plants under conditions of moderate selection pressure.
We demonstrate that recombinant viruses formed between a wild-type virus and a viral transgene can be isolated from transgenic plants under conditions of moderate to weak selection pressure. We inoculated cauliflower mosaic virus (CaMV) strain W260 to transgenic Nicotiana bigelovii plants that expressed a copy of CaMV gene VI derived from CaMV strain D4, a gene that determines systemic infection of solanaceous species, including N. bigelovii. Because W260 infects nontransformed N. bigelovii systemically, a recombinant virus formed between W260 and the D4 transgene would be expected to have little selective advantage over the wild-type W260 virus W260 was inoculated to approximately 100 plants each of nontransformed and transgenic N. bigelovii and it systemically infected nearly all of the plants. An analysis of viral DNA recovered from 23 transgenic plants infected with W260 revealed that 20 infections resulted from the systemic movement of the wild-type W260 virus, while a recombinant between W260 and the D4 transgene was detected in three of the infections. To determine the percentage of recovery of recombinant viruses under strong selection pressure, we inoculated approximately 100 nontransformed and 100 D4 gene VI transgenic plants with CaMV strain CM1841, a virus that is unable to infect nontransformed N. bigelovii. CM1841 infected 36% of the transgenic plants systemically, but none of the nontransformed controls. An analysis of 24 infected plants showed that a recombination event occurred in every plant, demonstrating that under strong selection conditions, the recovery of CaMV recombinants from transgenic plants can be very high.


Still lonely: Social adjustment of youth with and without social anxiety disorder following cognitive behavioral therapy.
Social experiences are an integral part of normative development for youth and social functioning difficulties are related to poor outcomes. Youth with anxiety disorders, and particularly social anxiety disorder, experience difficulties across many aspects of social functioning that may place them at risk for maladjustment. The goal of this paper was to compare social experiences of youth across anxiety diagnoses and examine whether treatment is helpful in improving social functioning. Ninety-two children (age 7-12 years; 58% male; 87.0% White) with a primary diagnosis of generalized anxiety disorder, separation anxiety disorder, and/or social anxiety disorder participated in cognitive behavioral therapy. At both pre- and post-treatment, children with social anxiety disorder self-reported greater loneliness than youth without social anxiety disorder, though levels of peer victimization and receipt of prosocial behavior were similar across groups. Parents reported greater social problems for youth with social anxiety disorder compared to those without social anxiety disorder. All youth experienced improved social functioning following treatment per child- and parent-reports. The results call for an increased focus on the social experiences of youth with anxiety disorders, and particularly loneliness, for children with social anxiety disorder. The results document ways that evidenced-based practice can improve social functioning for youth with anxiety disorders.


Measurement of human and mouse interleukin-12.
This unit describes functional assays for measurement of bioactive IL-12 and ELISAs for measurement of IL-12 protein. The functional assays are based on the ability of IL-12 to stimulate proliferation of PHA-activated T lymphoblasts ("PHA blasts"). The ELISAs are technically simpler to perform than the functional assays, but cannot distinguish bioactive from inactive cytokine.


A new free radical scavenger, edaravone, ameliorates oxidative liver damage due to ischemia-reperfusion in vitro and in vivo.
Ischemia-reperfusion injury causes oxidative stress producing reactive oxygen species, which is a serious problem linked to morbidity and mortality in liver surgery. We investigated the effects of edaravone, a new free radical scavenger, on liver oxidative stress in vitro and in vivo. We employed a hypoxia-reoxygenation model of primary cultured hepatocytes using an AnaeroPack (Mitsubishi Gas Chemical Co., Tokyo, Japan). Hepatocytes were exposed to 3 or 4 hours of hypoxia and then returned to oxygenation. We analyzed the time course changes of aspartate aminotransferase (AST), phosphatidylcholine hydroperoxide (PCOOH), and adenosine triphosphate (ATP) content in hepatocytes of edaravone-treated groups or nontreated groups after reoxygenation. Edaravone significantly attenuated the elevation of the AST level of the medium and hepatocellular PCOOH and preserved the hepatocellular ATP level. In vivo, male Sprague-Dawley rats were subjected to 45 minutes of hepatic ischemia and 120 minutes of reperfusion. The rats were intravenously injected with vehicle or edaravone (3 mg/kg or 10 mg/kg) before reperfusion and 1 hour after reperfusion. Serum AST levels and hepatic PCOOH and energy charge were significantly improved in both edaravone groups compared with control. In conclusion, edaravone has the ability to eliminate intra-hepatocellular superoxide species and attenuate oxidative liver damage in liver surgery.


Hunger tolerance and Leishmania in sandflies.
The sandfly Phlebotomus papatasi transmits Leishmania major, the agent of cutaneous leishmaniasis, in desert and savannah regions of the Old World, where seasonal stress of dehydration and heat reduces the quantity of sugar in plant leaves. Without essential sugar, only a few flies that feed on leaves can survive for long enough to deposit eggs and transmit Leishmania. Accordingly, selection for hunger tolerance may also select for pathogen susceptibility in flies. Here we provide evidence of a link between these advantageous and costly properties by testing the susceptibility of flies selected by sugar deprivation and of flies from irrigated and arid habitats.


[Risk of ileus, incisional and parastomal hernia after open and laparoscopic surgery for rectal cancer].