A novel star PEG-derived surface coating for specific cell adhesion.
In this study a novel approach for the coating and functionalization of substrates for cell culture and tissue engineering is presented. Glass, silicon, and titanium panes were coated with an ultrathin film (30 +/- 5 nm) of reactive star-shaped poly(ethylene glycol) prepolymers (Star PEG). Homogeneity of the films was checked by optical microscopy and scanning force microscopy. These coatings prevent unspecific protein adsorption as monitored by fluorescence microscopy and ellipsometry. In order to allow specific cell adhesion the films were modified with linear RGD peptides (gRGDsc) in different concentrations. After sterilization, fibroblast, SaOS, and human mesenchymal stem cells (hMSC) were seeded on these substrates. Cell adhesion, spreading, and survival was observed for up to 30 days on linear RGD peptide (gRGDsc)-modified coatings, whereas no cell adhesion could be detected on unmodified Star PEG layers. By variation of the RGD concentration within the film the amount of cells that became adhesive could be controlled. When differentiation conditions are used for cultivation of hMSCs the cells show the expression of osteogenic marker genes after 14 days which is comparable to cultivation on cell culture plastic. Thus, the Star PEG/RGD film did not negatively influence the differentiation process. The high flexibility of the system considering the incorporation of biologically active compounds opens a broad field of future experiments.


Heat-shock response in a molluscan cell line: characterization of the response and cloning of an inducible HSP70 cDNA.
Sublethal heat-shock of cells of the Bge (Biomphalaria glabrata embryonic) snail cell line resulted in increased or new expression of metabolically labeled polypeptides of approximately 21.5, 41, 70, and 74 kDa molecular mass. Regulation of this response appeared to be at the transcriptional level since a similar protein banding pattern was seen upon SDS-PAGE/fluorographic analysis of polypeptides produced by in vitro translation of total RNA from cells subjected to heat shock. Using a yeast (Saccharomyces cerevisiae) 70-kDa heat-shock protein (HSP70) probe to screen a cDNA library from heat-treated Bge cells, we isolated a full-length cDNA clone encoding a putative Bge HSP70. The cDNA was 2453 bp in length and contained an open reading frame of 1908 bp encoding a 636-amino-acid polypeptide with calculated molecular mass of 70,740 Da. Comparison of a conserved region of 209 amino acid residues revealed > 80% identity between the deduced amino acid sequence of Bge HSP70 and that of yeast (81%), the human blood fluke Schistosoma mansoni (for which B. glabrata serves as intermediate host) (81%), Drosophila (81%), human (84%), and the marine gastropod Aplysia californica (88%, 90%). In addition to the extensive sharing of sequence homology, the identification of several eukaryotic HSP70 signature sequences and an N-linked glycosylation site characteristic of cytoplasmic HSPs strongly support the identity of the Bge cDNA as encoding an authentic HSP70. Results of a Northern blot analysis, using Bge HSP70 clone-specific probes, indicated that gene expression was heat inducible and not constitutively expressed. This is the first reported sequence of an inducible HSP70 from cells originating from a freshwater gastropod and provides a first step in the development of a genetic transformation system for molluscs of medical importance.


Frequency-dependent fine structure in the frequency-following response: The byproduct of multiple generators.
The frequency-following response (FFR) is an auditory-evoked response recorded at the scalp that captures the spectrotemporal properties of tonal stimuli. Previous investigations report that the amplitude of the FFR fluctuates as a function of stimulus frequency, a phenomenon thought to reflect multiple neural generators phase-locking to the stimulus with different response latencies. When phase-locked responses are offset by different latencies, constructive and destructive phase interferences emerge in the volume-conducted signals, culminating in an attenuation or amplification of the scalp-recorded response in a frequency-specific manner. Borrowing from the literature on the audiogram and otoacoustic emissions (OAEs), we refer to this frequency-specific waxing and waning of the FFR amplitude as fine structure. While prior work on the human FFR was limited by small sets of stimulus frequencies, here, we provide the first systematic investigation of FFR fine structure using a broad stimulus set (90 + frequencies) that spanned the limits of human pitch perception. Consistent with predictions, the magnitude of the FFR response varied systematically as a function of stimulus frequency between 16.35 and 880 Hz. In our dataset, FFR high points (local maxima) emerged at ∼44, 87, 208, and 415 Hz with FFR valleys (local minima) emerging ∼62, 110, 311, and 448 Hz. To investigate whether these amplitude fluctuations are the result of multiple neural generators with distinct latencies, we created a theoretical model of the FFR that included six putative generators. Based on the extant literature on the sources of the FFR, our model adopted latencies characteristic of the cochlear microphonic (0 ms), cochlear nucleus (∼1.25 ms), superior olive (∼3.7 ms), and inferior colliculus (∼5 ms). In addition, we included two longer latency putative generators (∼13 ms, and ∼25 ms) reflective of the characteristic latencies of primary and non-primary auditory cortical structures. Our model revealed that the FFR fine structure observed between 16.35 and 880 Hz can be explained by the phase-interaction patterns created by six generators with relative latencies spaced between 0 and 25 ms. In addition, our model provides confirmatory evidence that both subcortical and cortical structures are activated by low-frequency (<100 Hz) tones, with the cortex being less sensitive to frequencies > 100 Hz. Collectively, these findings highlight (1) that the FFR is a composite response; (2) that the FFR at any given frequency can reflect activity from multiple generators; (3) that the fine-structure pattern between 16.35 and 880 Hz is the collective outcome of short- and long-latency generators; (4) that FFR fine structure is epiphenomenal in that it reflects how volume-conducted electrical potentials originating from different sources with different latencies interact at scalp locations, not how these different sources actually interact in the brain; and (5) that as a byproduct of these phase-interaction patterns low-amplitude responses will emerge at some frequencies, even when the underlying generators are fully functioning. We believe these findings call for a re-examination of how FFR amplitude is interpreted in both clinical and experimental contexts.


Sulfhydryl angiotensin-converting enzyme inhibition induces sustained reduction of systemic oxidative stress and improves the nitric oxide pathway in patients with essential hypertension.
Essential hypertension is associated with enhanced LDL oxidation and impaired endothelium-dependent vasodilation. The antioxidant status is linked to the nitric oxide (NO) pathway. Sulfhydryl angiotensin-converting enzyme (ACE) inhibitors inhibit oxidative stress and atherogenesis in experimental models; therefore we tested whether this beneficial antioxidant activity could be also clinically relevant in patients with essential hypertension. Plasma LDL oxidizability was investigated initially in untreated normocholesterolemic patients with moderate essential hypertension without clinically evident target organ damage (n = 96) and in control normotensive subjects (n = 46). Patients were then randomly assigned into two age- and sex-matched groups to receive the new sulfhydryl ACE inhibitor zofenopril (15 to 30 mg/d; n = 48) or enalapril (20 mg/d, n = 48). LDL oxidizability was evaluated (generation of malondialdehyde, MDA) and systemic oxidative stress was evaluated by isoprostanes (8-isoPGF2alpha). Asymmetrical dimethyl-L-arginine (ADMA), a competitive inhibitor of endothelial NO synthase, and plasma nitrite and nitrates (NOx) were also measured. LDL from hypertensive subjects had enhanced susceptibility to oxidation in vitro compared with that in control subjects (P <.05). Similarly, isoprostanes were significantly increased (P <.01) in hypertensive subjects versus control subjects. After 12-week treatment, MDA levels were significantly reduced by zofenopril (P <.05) but not enalapril treatment (P = not significant). Isoprostanes were normalized after zofenopril treatment (P <.03), whereas enalapril was ineffective. After treatment with both ACE inhibitors, plasma NOx concentrations were significantly reduced (P <.05). Similarly, hypertension increased ADMA concentration compared with the normotensive state, whereas ACE inihibition elicited a significant decrease. However, the reduction of ADMA concentration was significantly higher in patients receiving sulfhydryl ACE inhibition (P <.05 vs enalapril). The sulfhydryl ACE inhibitor zofenopril reduces oxidative stress and improves the NO pathway in patients with essential hypertension. If confirmed in a large multicenter clinical trial, our data suggest a possible vasculoprotective effect of the compound in retarding vascular dysfunction and atherogenesis that often develops rapidly in hypertensive patients.


Quantitative detection of type A staphylococcal enterotoxin by Laurell electroimmunodiffusion.
The detection of staphylococcal enterotoxin A by the quantitative technique of electroimmunodiffusion is described. High dilutions of type-specific rabbit antiserum were used in 1% agarose gels, 1 mm thick, and prepared in 0.05-mug barbital buffer, pH 8.6. Volumes of 10 muliters containing 1.5 to 10 ng of toxin were electrophoresed out of 4-mm diameter wells at 5 mA/cm width of gel. The precipitin cones formed were made visible by first immersing the agarose gels in 0.2 M NaCl and then overlaying the surface with the purified globulin fraction of sheep serum against rabbit globulin, followed by soaking of the gels in 1% aqueous cadmium acetate and staining with 0.1% thiazine red in 1% glacial acetic acid. Fully extended cones, 4 to 23 mm in length depending on toxin concentration and antiserum dilution, were developed in 2 to 5 h of electrophoresis, and visualization was achieved within 2 to 3 h. Because the method is qualitative, quantitative, simple, rapid, and sensitive, it offers a practical tool for the detection of small amounts of bacterial toxins in contaminated foods. The method should also qualify as a sensitive detection device in biochemical procedures which attempt to trace, detect, and identify biological substances in nanogram quantities, provided these substances are antigenic and capable of forming a precipitate with their specific antibodies.


Treatment of recurrent high grade astrocytoma; results of a systematic review of 1,415 patients.
The optimum treatment strategy for recurrent high grade glioma (anaplastic astrocytoma and glioblastoma multiforme) remains undefined. The objective of this report is to present a systematic analysis of the published clinical trial literature on the therapy of this disease. A study protocol was prospectively developed outlining the objectives and methods of the analysis such as; literature search strategy, eligibility criteria for published trials to be included, key data elements to be extracted, and a plan for statistical analysis. Variables of interest were, those describing key features of the study such as publication date and geographic location of reporting institution, characteristics of study population including functional status, details of patient treatment, treatment sequencing and data on clinical outcomes of interest, i.e. time to tumor progression and overall survival. Data were analyzed using SAS version 6.11. Summary statistics were calculated for the primary outcome variables. Data on recognized prognostic variables were recorded in order to adjust the analysis for these parameters. A total of 40 trials (36 non-randomized controlled trials; nRCTs and 4 randomized controlled trials: RCTs) were included in the analysis. Thirty-two of the eligible trials were chemotherapy trials while 7 were radiation therapy trials. One surgical trial met eligibility criteria. A total of 47 treatment arms were analysed which included 1,415 patients. Five chemotherapy groups were studied, i.e. interferons, nitrosoureas, platinums, platinums + nitrosoureas and others. The nitrosoureas were found to significantly extend time to tumor progression compared to all other drugs (26.9 weeks). The nitrosoureas and platinums appear to be the most active agents with regard to overall survival (over 32.0 weeks) as compared with the other drug categories. Patients treated with both a nitrosourea and a platinum compound showed the longest overall survival (40.0 weeks) although this was not significantly different from these drugs used as single agents. Average median survival for patients treated with radiation therapy was 44.7 weeks although selection bias makes these data difficult to compare with those derived from studies of chemotherapy. No definite conclusions can be made regarding surgical therapy in this setting due to limited data. The nitrosoureas and platinums, either as single agents or as combination chemotherapy, appear to be the most active agents in this disease although few, well designed chemotherapy trials are available for analysis. Due to the limitations of the available data on radiation therapy and surgery, as outlined in this report, additional, well designed clinical trials are needed to define the appropriate role for these modalities in the treatment of recurrent high grade glioma.


Combination of rTMS and treadmill training modulates corticomotor inhibition and improves walking in Parkinson disease: a randomized trial.
Repetitive transcranial magnetic stimulation (rTMS) of the brain has been shown to modulate cortical excitability. Combinations of rehabilitation therapies with rTMS might enhance the therapeutic effects. The purpose of this study was to investigate the effects of high-frequency rTMS followed by treadmill training on cortical inhibition and walking function in individuals with Parkinson disease (PD). A total of 20 patients with PD were randomized into an experimental group and a control group. Participants received rTMS (experimental group) or sham rTMS (control group) followed by treadmill training (30 minutes) for 12 sessions over 4 weeks. Repetitive TMS was applied at a 5-Hz frequency over the leg area of the motor cortex contralaterally to the more affected side for 6 minutes. Outcomes, including corticomotor inhibition and walking performance, were measured before and after training. . The results showed significant time effects on almost all corticomotor and functional variables. There are significant interaction effects between group and time of evaluation on the motor threshold, duration of the cortical silent period, and short interval intracortical inhibition of the contralateral hemisphere relatively to the more affected side as well as on the fast walking speed and timed up and go. The findings suggested that combination of rTMS and treadmill training enhances the effect of treadmill training on modulation of corticomotor inhibition and improvement of walking performance in those with PD.


How to study the neural mechanisms of multiple tasks.
Most biological and artificial neural systems are capable of completing multiple tasks. However, the neural mechanism by which multiple tasks are accomplished within the same system is largely unclear. We start by discussing how different tasks can be related, and methods to generate large sets of inter-related tasks to study how neural networks and animals perform multiple tasks. We then argue that there are mechanisms that emphasize either specialization or flexibility. We will review two such neural mechanisms underlying multiple tasks at the neuronal level (modularity and mixed selectivity), and discuss how different mechanisms can emerge depending on training methods in neural networks.


Antibiotic overproduction in Streptomyces coelicolor A3 2 mediated by phosphofructokinase deletion.
Streptomycetes are exploited for production of a wide range of secondary metabolites, and there is much interest in enhancing the level of production of these metabolites. Secondary metabolites are synthesized in dedicated biosynthetic routes, but precursors and co-factors are derived from the primary metabolism. High level production of antibiotics in streptomycetes therefore requires engineering of the primary metabolism. Here we demonstrate this by targeting a key enzyme in glycolysis, phosphofructokinase, leading to improved antibiotic production in Streptomyces coelicolor A3(2). Deletion of pfkA2 (SCO5426), one of three annotated pfkA homologues in S. coelicolor A3(2), resulted in a higher production of the pigmented antibiotics actinorhodin and undecylprodigiosin. The pfkA2 deletion strain had an increased carbon flux through the pentose phosphate pathway, as measured by (13)C metabolic flux analysis, establishing the ATP-dependent PfkA2 as a key player in determining the carbon flux distribution. The increased pentose phosphate pathway flux appeared largely because of accumulation of glucose 6-phosphate and fructose 6-phosphate, as experimentally observed in the mutant strain. Through genome-scale metabolic model simulations, we predicted that decreased phosphofructokinase activity leads to an increase in pentose phosphate pathway flux and in flux to pigmented antibiotics and pyruvate. Integrated analysis of gene expression data using a genome-scale metabolic model further revealed transcriptional changes in genes encoding redox co-factor-dependent enzymes as well as those encoding pentose phosphate pathway enzymes and enzymes involved in storage carbohydrate biosynthesis.


Nursing practice patterns: timing of insulin administration and glucose monitoring in the hospital.
The purpose of this study was to describe diabetes nursing practice patterns related to the timing of morning insulin administration, blood glucose monitoring, and meal intake for patients with type 2 diabetes and to report how frequently nurses were able to meet the expected standard of care. Observations were made of 50 incidents of morning diabetes care on medical-surgical units in an acute care setting. Timeliness of rapid-acting insulin administration and glucose monitoring in relation to the morning meal were evaluated, as were the subsequent prelunch glucose levels. Patients did not receive rapid-acting insulin with in the standard recommended time of 10 minutes premeal/postmeal 84% of the time, nor did they receive glucose monitoring within the recommended 30 minutes premeal 57% of the time. There was no significant relationship found between timely insulin administration and glucose monitoring. There were no significant relationships found between care activities and prelunch glucose control or glucose variability. Coordinating insulin administration, glucose monitoring, and meal delivery within the tight time frames required for rapid-acting insulin is a significant challenge not being met. Timeliness of diabetes nursing care is not the sole determining factor to good glucose control in hospitalized patients. Standards regarding timing of these activities need to be evaluated.


Random Sampling using k-vector.
This work introduces two new techniques for random number generation with any prescribed nonlinear distribution based on the k-vector methodology. The first approach is based on inverse transform sampling using the optimal k-vector to generate the samples by inverting the cumulative distribution. The second approach generates samples by performing random searches in a pre-generated large database previously built by massive inversion of the prescribed nonlinear distribution using the k-vector. Both methods are shown suitable for massive generation of random samples. Examples are provided to clarify these methodologies.


[Tuberculosis in general practice in Belgium : What's new?]
Tuberculosis is still an important disease in Belgium, mainly in big agglomerations. For this reason, knowledge of this entity by a general practitioner is very important. Genetic tests able to identify the species of Mycobacterium tuberculosis have been developed in recent years, but also identification of resistance to 1st and 2nd line treatment through genetic methods has been developed. The quick availability of results of this tests compared to the standard tests is a big advantage. Treatment of drug-susceptible tuberculosis hasn't changed recently. In contrast a lot has changed in the landscape of multidrug resistant tuberculosis with the development of 2nd line drugs and shortening of treatment duration in specific cases of multidrug resistant tuberculosis.


[Clinical observation on ankle-three-needle for treatment of nerve root pain of prolapse of lumbar intervertebral disc].
To observe the analgesic effect of ankle-three-needle on nerve root pain of prolapse of lumbar intervertebral disc. Three hundred and eighty cases were randomly divided into an ankle-three-needle group, a routine acupuncture group and a medication group. The ankle-three-needle group were treated with ankle-three-needle therapy, i. e. according to different prolapse segments, points Gentong No. 1, 2 and 3 were selected with lifting-kneading needle insertion method used and the needle was inserted along the skin; the routine acupuncture group were treated with acupuncture at Shenshu (BL 23), Qihaishu (BL 24) and Jiaji (EX-B 2) of the prolapse corresponding segment, and Ciliao (BL 32), etc. with uniform reinforcing-reducing manipulation; the medication group were treated with routine buttock intramuscular injection of aspirin-DL-lysine plus saline. The time inducing analgesia was 6 min in the ankle-three-needle group, 27 min in the routine acupuncture group and 18 min in the medication group. The effect-lasting time was 24.48 h in the ankle-three-needle group, 8.93 h in the routine acupuncture group and 6.36 h in the medication group, with a significant difference as the ankle-three-needle group compared with both the routine acupuncture group and the medication group, but with no significant difference between the routine acupuncture group and the medication group. After treatment, there were very significant differences in change tendency of the analgesic score among the three groups at all the time points (P < 0.01). And there was very significant differences in the changing tendency of straight-leg raising test among the three groups at 0.5 h, 1 h, 24 h and 48 h (P < 0.001). Ankle-three-needle has obvious therapeutic effect on nerve root pain induced by prolapse of lumbar intervertebral disc.


Vibrational modes of ubiquinone in cytochrome bo(3) from Escherichia coli identified by Fourier transform infrared difference spectroscopy and specific (13)C labeling.
In this study we present the infrared spectroscopic characterization of the bound ubiquinone in cytochrome bo(3) from Escherichia coli. Electrochemically induced Fourier transform infrared (FTIR) difference spectra of DeltaUbiA (an oxidase devoid of bound ubiquinone) and DeltaUbiA reconstituted with ubiquinone 2 and with isotopically labeled ubiquinone 2, where (13)C was introduced either at the 1- or at the 4-position of the ring (C=O groups), have been obtained. The vibrational modes of the quinone bound to the discussed high-affinity binding site (Q(H)) are compared to those from the synthetic quinones in solution, leading to the assignment of the C=O modes to a split signal at 1658/1668 cm(-)(1), with both carbonyls similarly contributing. The FTIR spectra of DeltaUbiA reconstituted with the labeled quinones indicate an essentially symmetrical and weak hydrogen bonding of the two C=O groups from the neutral quinone with the protein and distinct conformations of the 2- and 3-methoxy groups. Perturbations of the vibrational modes of the 5-methyl side groups are discussed for a signal at 1452 cm(-)(1). Only negligible shifts of the aromatic ring modes can be reported for the reduced and the protonated form of the quinone. Alterations of the protein upon quinone binding are reflected in the electrochemically induced FTIR difference spectra. In particular, difference signals at 1640-1633 cm(-)(1) and 1700-1670 cm(-)(1) indicate variations of beta-sheet secondary structure elements and loops, bands at 1706 and 1678 cm(-)(1) are tentatively attributed to individual amino acids, and a difference signal a 1540 cm(-)(1) is discussed to reflect an influence on C=C modes of the porphyrin ring or on deprotonated propionate groups of the hemes. Further tentative assignments are presented and discussed. The (13)C labeling experiments allow the assignment of the vibrational modes of a bound ubiquinone 8 in the electrochemically induced FTIR difference spectra of wild-type bo(3).


Complete canthi removal reveals that forces from the amnioserosa alone are sufficient to drive dorsal closure in Drosophila.
Drosophila's dorsal closure provides an excellent model system with which to analyze biomechanical processes during morphogenesis. During native closure, the amnioserosa, flanked by two lateral epidermal sheets, forms an eye-shaped opening with canthi at each corner. The dynamics of amnioserosa cells and actomyosin purse strings in the leading edges of epidermal cells promote closure, whereas the bulk of the lateral epidermis opposes closure. Canthi maintain purse string curvature (necessary for their dorsalward forces), and zipping at the canthi shortens leading edges, ensuring a continuous epithelium at closure completion. We investigated the requirement for intact canthi during closure with laser dissection approaches. Dissection of one or both canthi resulted in tissue recoil and flattening of each purse string. After recoil and a temporary pause, closure resumed at approximately native rates until slowing near the completion of closure. Thus the amnioserosa alone can drive closure after dissection of one or both canthi, requiring neither substantial purse string curvature nor zipping during the bulk of closure. How the embryo coordinates multiple, large forces (each of which is orders of magnitude greater than the net force) during native closure and is also resilient to multiple perturbations are key extant questions.


Hepatitis C virus-infected women have a higher risk of advanced fibrosis and graft loss after liver transplantation than men.
In natural history studies of hepatitis C virus (HCV) infection, women have a lower risk of disease progression to cirrhosis. Whether female sex influences outcomes of HCV in the posttransplantation setting is unknown. All patients transplanted for HCV-related liver disease from 2002-2007 at five United States transplantation centers were included. The primary outcome was development of advanced disease, defined as biopsy-proven bridging fibrosis or cirrhosis. Secondary outcomes included death, graft loss, and graft loss with advanced recurrent disease. A total of 1,264 patients were followed for a median of 3 years (interquartile range, 1.8-4.7), 304 (24%) of whom were women. The cumulative rate of advanced disease at 3 years was 38% for women and 33% for men (P=0.31), but after adjustment for recipient age, donor age, donor anti-HCV positivity, posttransplantation HCV treatment, cytomegalovirus infection and center, female sex was an independent predictor of advanced recurrent disease (hazard ratio [HR], 1.31; 95% confidence interval [CI], 1.02-1.70; P=0.04). Among women, older donor age and treated acute rejection were the primary predictors of advanced disease. The unadjusted cumulative 3-year rates of patient and graft survival were numerically lower in women (75% and 74%, respectively) than men (80% and 78%, respectively), and in multivariable analyses, female sex was an independent predictor for death (HR, 1.30; 95% CI, 1.01-1.67; P=0.04) and graft loss (HR, 1.31; 95% CI, 1.02-1.67; P=0.03). Female sex represents an underrecognized risk factor for advanced recurrent HCV disease and graft loss. Further studies are needed to determine whether modification of donor factors, immunosuppression, and posttransplantation therapeutics can equalize HCV-specific outcomes in women and men.


Synthesis and Antibacterial Evaluation of Novel 3-Substituted Ocotillol-Type Derivatives as Leads.
Due to the rapidly growing bacterial antibiotic-resistance and the scarcity of novel agents in development, bacterial infection is still a global problem. Therefore, new types of antibacterial agents, which are effective both alone and in combination with traditional antibiotics, are urgently needed. In this paper, a series of antibacterial ocotillol-type C-24 epimers modified from natural 20(S)-protopanaxadiol were synthesized and evaluated for their antibacterial activity. According to the screening results of Gram-positive bacteria (B. subtilis 168 and MRSA USA300) and Gram-negative bacteria (P. aer PAO1 and A. baum ATCC19606) in vitro, the derivatives exhibited good antibacterial activity, particularly against Gram-positive bacteria with an minimum inhibitory concentrations (MIC) value of 2-16 µg/mL. The subsequent synergistic antibacterial assay showed that derivatives 5c and 6c enhanced the susceptibility of B. subtilis 168 and MRSA USA300 to chloramphenicol (CHL) and kanamycin (KAN) (FICI < 0.5). Our data showed that ocotillol-type derivatives with long-chain amino acid substituents at C-3 were good leads against antibiotic-resistant pathogens MRSA USA300, which could improve the ability of KAN and CHL to exhibit antibacterial activity at much lower concentrations with reduced toxicity.


Emergence of gentamicin- and carbenicillin-resistant Pseudomonas aeruginosa in a hospital environment.
Strains of Pseudomonas aeruginosa resistant to either gentamicin or carbenicillin have been noted since their introduction into clinical use. During a 6-month period, twice-weekly cultures were obtained from all patients treated with either gentamicin or carbenicillin and from all patients with a positive culture for P. aeruginosa. Susceptibility testing to gentamicin and carbenicillin and pyocine typing were performed on all isolates. Organisms with a minimal inhibitory concentration greater than 12.5 mug of gentamicin per ml or greater than 100 mug of carbenicillin per ml were defined as resistant. P. aeruginosa was cultured from 238 patients. One patient was initially infected with a gentamicin-resistant isolate. In 11 other patients, serial cultures revealed the emergence of resistance to gentamicin. All but one of these resistant isolates occurred in patients treated with gentamicin. In eight instances the pyocine and/or serological types before and after the change in sensitivity pattern were the same. Gentamicin-resistant P. aeruginosa emerged significantly more often in patients treated with gentamicin than in those who did not receive gentamicin. Carbenicillin-resistant P. aeruginosa emerged in four of 14 patients treated with carbenicillin. Seventeen of the 238 patients were infected de novo with carbenicillin-resistant P. aeruginosa. Carbenicillin-resistant P. aeruginosa emerged significantly more often in patients treated with carbenicillin than in those who did not receive carbenicillin. No evidence was found for in-hospital spread of resistant P. aeruginosa.


C-banding and fluorescence in situ hybridization studies of the wheat-alien hybrid 'Agrotana'.
'Agrotana', a wheat-alien hybrid (2n = 56), is a potential source of resistance to common root rot, stem rust, wheat streak mosaic virus, and the wheat curl mite. However, the origin of 'Agrotana', reported to be durum wheat x Agropyron trichophorum (pubescent wheatgrass), is uncertain. The objective of this investigation was to determine the chromosome constitution of 'Agrotana' using C-banding and fluorescence in situ hybridization techniques. The F1 hybrid of 'Agrotana' x 'Chinese Spring' wheat showed 7 I + 21 II in 14.9% of the pollen mother cells, evidence of the presence of the A, B, and D genomes in 'Agrotana'. The hybrid had 16 heavily C-banded chromosomes, namely 4A, and 1-7B of wheat, and a translocation that probably involved wheat chromosomes 2A and 2D. In situ hybridization using biotinylated genomic DNA of Ag. trichophorum cv. Greenleaf blocked with CS DNA failed to identify the alien chromosomes in 'Agrotana', indicating that the alien chromosomes were not likely derived from pubescent wheatgrass. In situ hybridization using labelled wheat genomic DNA blocked with 'Agrotana' DNA revealed that 'Agrotana' had 40 wheat, 14 alien, and 2 (a pair) wheat-alien translocated chromosomes. There was no homology between wheat and the alien chromosomes or chromosome segments involved in the wheat-alien recombinant. Two of the seven pairs of alien chromosomes were homoeologous to each other. The ability to identify alien chromatin in wheat using labelled wheat DNA instead of labelled alien DNA will be particularly useful in chromosome engineering of wheat germplasms having alien chromatin of unknown origin.


Evaluation of the safety performance of highway alignments based on fault tree analysis and safety boundaries.
To overcome the limitations of previous highway alignment safety evaluation methods, this article presents a highway alignment safety evaluation method based on fault tree analysis (FTA) and the characteristics of vehicle safety boundaries, within the framework of dynamic modeling of the driver-vehicle-road system. Approaches for categorizing the vehicle failure modes while driving on highways and the corresponding safety boundaries were comprehensively investigated based on vehicle system dynamics theory. Then, an overall crash probability model was formulated based on FTA considering the risks of 3 failure modes: losing steering capability, losing track-holding capability, and rear-end collision. The proposed method was implemented on a highway segment between Bengbu and Nanjing in China. A driver-vehicle-road multibody dynamics model was developed based on the 3D alignments of the Bengbu to Nanjing section of Ning-Luo expressway using Carsim, and the dynamics indices, such as sideslip angle and, yaw rate were obtained. Then, the average crash probability of each road section was calculated with a fixed-length method. Finally, the average crash probability was validated against the crash frequency per kilometer to demonstrate the accuracy of the proposed method. The results of the regression analysis and correlation analysis indicated good consistency between the results of the safety evaluation and the crash data and that it outperformed the safety evaluation methods used in previous studies. The proposed method has the potential to be used in practical engineering applications to identify crash-prone locations and alignment deficiencies on highways in the planning and design phases, as well as those in service.


Discovery and characterization of orally bioavailable 4-chloro-6-fluoroisophthalamides as covalent PPARG inverse-agonists.
PPAR gamma (PPARG) is a ligand activated transcription factor that regulates genes involved in inflammation, bone biology, lipid homeostasis, as well as a master regulator of adipogenesis and a potential lineage driver of luminal bladder cancer. While PPARG agonists lead to transcriptional activation of canonical target genes, inverse agonists have the opposite effect through inducing a transcriptionally repressive complex leading to repression of canonical target gene expression. While many agonists have been described and tested clinically, inverse agonists offer an underexplored avenue to modulate PPARG biology in vivo. Current inverse agonists lack favorable in vivo properties; herein we describe the discovery and characterization of a series of orally bioavailable 4-chloro-6-fluoroisophthalamides as covalent PPARG inverse-agonists, BAY-5516, BAY-5094, and BAY-9683. Structural studies of this series revealed distinct pre- and post-covalent binding positions, which led to the hypothesis that interactions in the pre-covalent conformation are primarily responsible for driving affinity, while interactions in the post-covalent conformation are more responsible for cellular functional effects by enhancing PPARG interactions with its corepressors. The need to simultaneously optimize for two distinct states may partially explain the steep SAR observed. Exquisite selectivity was achieved over related nuclear receptors in the subfamily due in part to a covalent warhead with low reactivity through an SNAr mechanism in addition to the specificity gained through covalent binding to a reactive cysteine uniquely positioned within the PPARG LBD. BAY-5516, BAY-5094, and BAY-9683 lead to pharmacodynamic regulation of PPARG target gene expression in vivo comparable to known inverse agonist SR10221 and represent new tools for future in vivo studies to explore their potential utility for treatment of disorders of hyperactivated PPARG including luminal bladder cancer and other disorders.


Osteomyelitis in patients with systemic lupus erythematosus.
To investigate the clinical profile of and the risk factors for osteomyelitis in patients with systemic lupus erythematosus (SLE). We reviewed 11 consecutive cases of patients with SLE who had also had osteomyelitis between 1981 and 2001 at a medical center in Taiwan, with special attention to predisposing factors, clinical features, laboratory values, and outcomes. The mean age at diagnosis of osteomyelitis was 34.5 +/- 22.0 years and the ratio of females to males was 9:2. The typical initial manifestations were nonspecific focal pain (82%) and fever (64%). The most commonly affected sites were the long bones (6 cases, 54%), followed by the vertebrae (4 cases, 36%). Salmonella (5 cases, 45%) and Staphylococcus aureus (4 cases, 36%) were the major causative organisms. Interestingly, once long bones had become involved, 5 of 6 (83%) isolates proved to be Salmonella, and for vertebral osteomyelitis, 3 of 4 (75%) isolates proved to be S. aureus. Predisposing factors include an active status of SLE (SLEDAI score >/= 4, 100%), coexistent underlying systemic disease (91%), chronic renal disease (82%), and intensified immunosuppressive agent usage (82%). Laboratory values either reflected an acute phase reaction that would be expected in an infection, such as a raised C-reactive protein (100%) and neutrophilia (55%), or reflected features consistent with active lupus disease. Four patients had longterm motor deficits and another patient died. Poor prognostic factors include delayed diagnosis, vertebral involvement, artificial implants in bones, and chronic carrier status. In patients with SLE who present with local osteoarticular pain, particularly those whose disease is active and who also have chronic renal disease and were taking intensified immunosuppressive agents, osteomyelitis must be considered seriously. Salmonella should be considered as a potential contributing pathogen for long bone osteomyelitis and S. aureus should be considered for cases of vertebral osteomyelitis when conducting empirical antimicrobial therapy. Early recognition and treatment is essential to avoid longterm sequelae or death.


The current acceptance, accessibility and recognition of Chinese and Ayurvedic medicine in the United States in the public, governmental, and industrial sectors.
To assess the current level of acceptance in the United States of complementary and alternative medicine, recent research into the prevalence, acceptance, accessibility, and recognition of complementary and alternative therapies were reviewed. Several signs point to an increasing acceptance of complementary and alternative medicine in the United States; the use of complementary and alternative medicine is significantly increasing, many aspects of Chinese medicine and Ayurveda are becoming mainstream, practitioners in the United States are beginning to be licensed, and insurance companies are beginning to cover some complementary and alternative therapies. Remaining challenges to true acceptance include the restrictive Western mindset, the absence of published studies, a lack of consistent manufacturing processes and quality standards, and a fear of adulteration. Although the field still faces many challenges, alternative and complementary medicine, including Chinese medicine and Ayurvedic medicine, is becoming more accepted and accessible in the United States.


The analysis of densitometric and geometric parameters of bilateral proximal phalanges in horses with the use of peripheral quantitative computed tompgraphy.
Proximal phalanges in horses are among bones that are most prone to injuries. So far, the detailed analysis of densitometric and geometric parameters of both front legs proximal phalanges in horses has not been investigated. The aim of this study was to compare the densitometric and geometric parameters between proximal phalanges in equine both front legs with the use of peripheral quantitative computed tomography (pQCT). The study material comprised isolated both front legs proximal phalanges derived from 22 horses. The structure analysis of the proximal phalanges was conducted with the pQCT. The following bone parameters were determined: bone mineral content, volumetric bone mineral density, total bone area, trabecular area, cortical area, cortical thickness, periosteal circumference, endocortical circumference, Strength Strain Index. Tomographic analysis of proximal phalanges was conducted at three levels: at 15%, 50% and 85% of the bone length. The statistical analysis showed that both the densitometric and geometric parameters of the bone at 50% and 85% of its length, did not present any statistically significant differences for the left or right proximal phalanges of the forelimb. At the same time, all examined parameters measured at 15% of the bone length, in the vicinity of the proximal metaphysis revealed significant statistical differences between both front legs proximal phalanges. The proximal phalanx parameters in the forelimbs are significantly different for the left and right proximal phalanx at 15% of the length and they indicate higher Strength Strain Index of the left bone in this location. The densitometric and geometric parameters of the bone at 50% and 85% of its length, did not present any statistically significant differences for the left or right proximal phalanges of the left and right forelimbs. The most serious changes caused by asymmetrical load of the thoracic limbs in horses occur near the proximal metaphysis, where the spongious substance is most abundant. This may happen because the metabolism of the spongious bone tissue is eight times faster compared to the compact bone tissue. Thus, any changes, including those caused by asymmetrical strain exerted on the right and left thoracic limbs, are the earliest to be observed.


Determining similarity of scientific entities in annotation datasets.
Linked Open Data initiatives have made available a diversity of scientific collections where scientists have annotated entities in the datasets with controlled vocabulary terms from ontologies. Annotations encode scientific knowledge, which is captured in annotation datasets. Determining relatedness between annotated entities becomes a building block for pattern mining, e.g. identifying drug-drug relationships may depend on the similarity of the targets that interact with each drug. A diversity of similarity measures has been proposed in the literature to compute relatedness between a pair of entities. Each measure exploits some knowledge including the name, function, relationships with other entities, taxonomic neighborhood and semantic knowledge. We propose a novel general-purpose annotation similarity measure called 'AnnSim' that measures the relatedness between two entities based on the similarity of their annotations. We model AnnSim as a 1-1 maximum weight bipartite match and exploit properties of existing solvers to provide an efficient solution. We empirically study the performance of AnnSim on real-world datasets of drugs and disease associations from clinical trials and relationships between drugs and (genomic) targets. Using baselines that include a variety of measures, we identify where AnnSim can provide a deeper understanding of the semantics underlying the relatedness of a pair of entities or where it could lead to predicting new links or identifying potential novel patterns. Although AnnSim does not exploit knowledge or properties of a particular domain, its performance compares well with a variety of state-of-the-art domain-specific measures. Database URL: http://www.yeastgenome.org/


Error Detection-Based Model to Assess Educational Outcomes in Crisis Resource Management Training: A Pilot Study.
Otolaryngology-head and neck surgery (OTL-HNS) residents face a variety of difficult, high-stress situations, which may occur early in their training. Since these events occur infrequently, simulation-based learning has become an important part of residents' training and is already well established in fields such as anesthesia and emergency medicine. In the domain of OTL-HNS, it is gradually gaining in popularity. Crisis Resource Management (CRM), a program adapted from the aviation industry, aims to improve outcomes of crisis situations by attempting to mitigate human errors. Some examples of CRM principles include cultivating situational awareness; promoting proper use of available resources; and improving rapid decision making, particularly in high-acuity, low-frequency clinical situations. Our pilot project sought to integrate CRM principles into an airway simulation course for OTL-HNS residents, but most important, it evaluated whether learning objectives were met, through use of a novel error identification model.


Implementing High-Quality Primary Care Through a Health Equity Lens.
The COVID-19 pandemic highlighted the importance of centering health equity in future health system and primary care reforms. Strengthening primary care will be needed to correct the longstanding history of mistreatment of First Nations/Indigenous and racialized people, exclusion of health care workers of color, and health care access and outcome inequities further magnified by the COVID-19 pandemic. The National Academies of Sciences, Engineering, and Medicine (NASEM) released a report on Implementing High-Quality Primary Care: Rebuilding the Foundation of Health Care, that provided a framework for defining high-quality primary care and proposed 5 recommendations for implementing that definition. Using the report's framework, we identified health equity challenges and opportunities with examples from primary care systems in the United States and Canada. We are poised to reinvigorate primary care because the recent pandemic and the attention to continued racialized police violence sparked renewed conversations and collaborations around equity, diversity, inclusion, and health equity that have been long overdue. The time to transition those conversations to actionable items to improve the health of patients, families, and communities is now.Appeared as Annals "Online First" article.


Aging and development in social insects with emphasis on the honey bee, Apis mellifera L.
Honey bee colonies typically consist of about 20-40 thousand workers, zero to few thousand males (drones), depending on the time of year, and a single queen, the mother of the colony. Workers typically live 3-6 weeks during the spring and summer and can live about 4months during the winter. Queens are longer lived. Anecdotes of queens living 2-3years are not unusual, though they normally live less than a year in commercial hives. Little is known about the life span of drones. Queens develop from fertilized eggs that are not different from the eggs that develop into workers. Queens are, however, twice as large, have specialized anatomy, live much longer, and develop faster from egg to adult. All of these differences are derived from differences in larval rearing environment, primarily nutrition. The developmental trajectory of a female larva from worker into a queen can be determined as late as the third day of larval development, after this time the developmental pathway is fixed for a worker phenotype. The total time of larval development is only 5-6 days, therefore, just 2-3 days of differential feeding can lead to profound differences in development, and longevity. Workers undergo age development after they become adults. Workers usually initiate foraging behavior when they are 2-3 weeks old. The age at which a worker initiates foraging is a strong determinant of her length of life. This is presumed to be a result of the hazards of foraging, but natural senescence also occurs. Some bees remain in the nest and are never observed to forage, thereby outliving their forager sisters. Corresponding to this behavioral development are changes in the sizes of glands and the production of glandular products, increases in biogenic amine titers within the brain, an increase in the volume of specific regions of the brain, and changes in the neural system that affect perception of stimuli, and learning and memory. These age-related changes in behavior are regulated by intrinsic and extrinsic factors. Genetic variation has been demonstrated for many of these life history and behavioral traits. Selection and genome mapping studies have demonstrated relationships between the neural system, behavior, and life history traits.


High intracolonic acetaldehyde values produced by a bacteriocolonic pathway for ethanol oxidation in piglets.
Human colonic contents and many colonic microbes produce considerable amounts of acetaldehyde from ethanol in vitro. To examine in piglets if acetaldehyde is produced in the colon also in vivo, and if so, what is the fate of intracolonically formed acetaldehyde. Seventeen native, non-fasted female piglets (20-25 kg) were used. Six piglets received either 1.5 g/kg bw or 2.5 g/kg bw of ethanol intravenously. In seven piglets, 0.7 g or 1.75 g of ethanol/kg bw was administered intravenously, followed by a subsequent intragastric ethanol infusion of 1.8 g/kg bw and 4.5 g/kg bw, respectively. The samples of colonic contents for the assessment of ethanol and acetaldehyde concentrations were obtained up to seven hours. In four additional piglets, the intracolonic values of ethanol, acetaldehyde, and acetate were observed for 60 minutes after an intracolonic infusion of acetaldehyde solution. A raised intracolonic, endogenous acetaldehyde concentration (mean (SEM); 36 (9) microM) was found in all piglets before ethanol infusion. After the infusion of ethanol, intracolonic ethanol and acetaldehyde values increased in parallel, reaching the peak values 57 (4) mM of ethanol and 271 (20) microM of acetaldehyde in the group that received the highest dose of ethanol. A positive correlation (r = 0.45; p < 0.001) was found between intracolonic ethanol and acetaldehyde values. Acetaldehyde administered intracolonically was mainly metabolised to acetate but also to ethanol in the colon. Significant endogenous intracolonic acetaldehyde values can be found in the normal porcine colon. Furthermore, our results suggest the existence of a bacteriocolonic pathway for ethanol oxidation. Increased amounts of acetaldehyde are formed intracolonically from ingested ethanol by this pathway.


Amiloride-sensitive sodium-hydrogen exchange in osmotically shrunken rabbit red blood cells.
As part of a detailed study of cell volume regulation in high-potassium mammalian erythrocytes, we have characterized ouabain-insensitive sodium transport in normal and osmotically shrunken rabbit red cells. In cells of normal volume and physiological pH, there is no amiloride-inhibited component of the sodium efflux (into either sodium-containing or sodium-free media). Osmotic shrinkage activates an amiloride-sensitive (50% inhibitory concentration = 10(-5) M) sodium transport system that can catalyze net sodium movement in either direction. This system appears to be distinct from the sodium-sodium (sodium-lithium) counter-transporter that operates in cells of normal volume. Replacement of chloride with acetate does not inhibit the sodium flux, but replacement with either nitrate or thiocyanate is inhibitory. An inward sodium gradient in shrunken cells induces a net uphill efflux of acid equivalents, indicating that the sodium transport is a sodium-hydrogen exchange. However, a sevenfold inward gradient of hydrogen ions (pHo = 6.4; pHi = 7.2) does not stimulate net sodium efflux in shrunken cells. This suggests that the extracellular affinity of the transport site for hydrogen ions is high, and that there is an extracellular noncompetitive inhibitory site for proton binding. Bilateral pH reduction stimulates an amiloride-inhibitable sodium flux in cells of normal volume; this indicates that, as has been found in kidney, brain, and lymphocytes, there is an intracellular protonation site that can activate the transport. Shrinkage of the cells shifts the pH dependence of the transport, suggesting that part of the signal for the osmotic activation of the transport is a shift in the pKa of this modifier site.


Computer-tailored intervention increases colorectal cancer screening among low-income African Americans in primary care: Results of a randomized trial.
Although African Americans have the highest colorectal cancer (CRC) incidence and mortality rates of any racial group, their screening rates remain low. This randomized controlled trial compared efficacy of two clinic-based interventions for increasing CRC screening among African American primary care patients. African American patients from 11 clinics who were not current with CRC screening were randomized to receive a computer-tailored intervention (n = 335) or a non-tailored brochure (n = 358) designed to promote adherence to CRC screening. Interventions were delivered in clinic immediately prior to a provider visit. Univariate and multivariable logistic regression models analyzed predictors of screening test completion. Moderators and mediators were determined using multivariable linear and logistic regression analyses. Significant effects of the computer-tailored intervention were observed for completion of a stool blood test (SBT) and completion of any CRC screening test (SBT or colonoscopy). The colonoscopy screening rate was higher among those receiving the computer-tailored intervention group compared to the nontailored brochure but the difference was not significant. Predictors of SBT completion were: receipt of the computer-tailored intervention; being seen at a Veterans Affairs Medical Center clinic; baseline stage of adoption; and reason for visit. Mediators of intervention effects were changes in perceived SBT barriers, changes in perceived colonoscopy benefits, changes in CRC knowledge, and patient-provider discussion. Moderators of intervention effects were age, employment, and family/friend recommendation of screening. This one-time computer-tailored intervention significantly improved CRC screening rates among low-income African American patients. This finding was largely driven by increasing SBT but the impact of the intervention on colonoscopy screening was strong. Implementation of a CRC screening quality improvement program in the VA site that included provision of stool blood test kits and follow-up likely contributed to the strong intervention effect observed at that site. The trial is registered at ClinicalTrials.gov as NCT00672828.


Lack of effect of small high-dose volumes on the dose-response relationship for the development of fibrosis in distant parts of the ipsilateral lung in mini-pigs.
Multi-field radiation therapy for intrathoracic tumours results in a heterogeneous dose distribution in lung tissue. This study investigated whether irradiation of small lung volumes with high fibrogenic doses affects the dose-response relationship for development of fibrosis in distant parts of the ipsilateral lung of mini-pigs. The whole right lung of 26 'Mini-Lewe' pigs was irradiated with homogeneous doses of between 25 Gy and 40 Gy given in five equal fractions using opposing anterior-posterior portals and a linear accelerator. Another 32 animals were irradiated with a constant dose of 35 Gy to a small house-shaped high-dose field (base 3.0 cm, height 4 cm) located 3 cm caudolateral to the right hilus, while the surrounding right lung received either no irradiation or homogeneous doses of between 20 Gy and 30 Gy. The radiation fields were simulated and port films were obtained for each of the 10 fields in all pigs. Fibrosis was quantified 9 months after irradiation by determination of the hydroxyproline (HP) content of the 32 high-dose volumes and in the lung apex and the basolateral lung of all 58 pigs. Based on the reference value for the HP-ratio, i.e. the HP-concentration of the right lung over the left lung, obtained in 12 unirradiated control animals, the experimental results were converted into quantal data for probit analysis, a responder being an animal with an HP-ratio > 1.33. A dose-response relationship for the HP-ratio was obtained in the different lung sites and irradiation groups. For a given dose level the mean HP-ratios and response rates did not differ systematically between the lung apex and the basolateral lung. Probit analysis of the pooled data produced ED50 values of 21.8 Gy (95% CI 12-37) for irradiation without a high-dose volume and 25.9 Gy (24-28) for irradiation with a high-dose volume. These values are not significantly different. The results from both irradiation groups could be well fitted by a common dose-response curve with an ED50 value of 26.1 Gy. Unexpectedly, the response rates in the high-dose volume increased with increasing dose to the surrounding right lung. Analysis of the port films provided an explanation for this finding: inaccuracies in daily field positioning. When this error was corrected for by use of the mean dose to the high-dose volume, a dose-response curve with an ED50 of 25.2 Gy (22-29) was determined for the high-dose volume. The results of the study indicate that the irradiation of a small lung volume with high fibrogenic doses does not affect the dose-response relationship for development of fibrosis in distant parts of the ipsilateral lung.


Simple formulas for screening abnormal blood pressure in children and adolescents.
Childhood hypertension has been extensively focused on in the past decades because of its increasing incidence, which is related to physicians' awareness and the increasing number of obese children. Age, gender, and body size are the main determinants of blood pressure in children. The revised childhood blood pressure tables of the National High Blood Pressure Education Program are a prerequisite for classification of childhood hypertension. Although these tables provide a reasonable basis, they are intricate and height percentile is needed for final diagnosis. Many attempts have been done to decrease such complexity. We present new formulas that are concise and memorable, and will help physicians to screen prehypertensive and hypertensive pediatric patients.


Development and Evaluation of an Approach to Using Value of Information Analyses for Real-Time Prioritization Decisions Within SWOG, a Large Cancer Clinical Trials Cooperative Group.
Value of information (VOI) analyses can align research with areas with the greatest potential impact on patient outcome, but questions remain concerning the feasibility and acceptability of these approaches to inform prioritization decisions. Our objective was to develop a process for calculating VOI in "real time" to inform trial funding decisions within SWOG, a large cancer clinical trials group. We developed an efficient and scalable VOI modeling approach using a selected sample of 9 randomized phase II/III trial proposals from the Breast, Gastrointestinal, and Genitourinary Disease Committees reviewed by SWOG's leadership between 2008 and 2013. There was bidirectional communication between SWOG investigators and the research team throughout the modeling development. Partial expected value of sample information for the treatment effect evaluated by the proposed trial's primary endpoint was calculated using Monte Carlo simulation. We derived prior uncertainty in the treatment effect estimate from the sample size calculations. Our process was feasible for 8 of 9 trial proposals and efficient: the time required of 1 researcher was <1 week per proposal. We accommodated stakeholder input primarily by deconstructing VOI metrics into expected health benefits and incremental healthcare costs and assuming treatment decisions within our simulations were based on health benefits. Following customization, feedback from over 200 SWOG members was positive regarding the overall VOI framework, specific retrospective results, and potential for VOI analyses to inform future trial proposal evaluations. We developed an efficient and customized process to calculate the expected VOI of cancer clinical trials that is feasible for use in decision making and acceptable to investigators. Prospective use and evaluation of this approach is currently underway within SWOG.


Development stage distribution as a proxy for feeding success and growth for first feeding Norwegian spring spawning herring larvae.
The estimation of growth rates in young herring larvae (Clupea harengus) in the field can be difficult because the primary increments in the otoliths may not be discernible or formed at a daily level. Likewise, the estimation of mortality rates of fish larvae in the field is very difficult to achieve, especially in a rigorous quantitative manner. In this study, the authors suggest the use of a stage-based proxy of feeding success, growth and potential survival or mortality risk of field-caught larvae. The stage-based proxy is derived based on observations from previous laboratory studies where larvae successfully completing start-feeding on external food sources will advance through the early development stages, whereas those that do not (unsuccessful larvae) remain and accumulate in the development stage preceding first feeding. The relative occurrence of larvae in the early development stages is therefore expected to reflect feeding conditions of the larvae, with higher ratios of unsuccessful larvae indicative of poor feeding success and higher mortality risk. Using field data on Norwegian spring spawning herring, the authors document that the relative occurrence of larvae in the late non-feeding stage is significantly higher at lower average zooplankton concentrations, in line with the predictions of the authors that this novel approach of using a stage-based proxy could be a useful indication of feeding success, growth and mortality in the field. Further, there was a significant interaction effect with ambient temperature, with the ratio being higher at low zooplankton concentrations at higher temperatures. This study also suggests that these findings are not population specific as the same accumulation of non-feeding larvae in the late non-feeding stage was observed in laboratory-reared larvae of both autumn and spring spawning herring populations.


Sacral resection. Operative technique and outcome.
Tumors involving the sacrum are difficult to treat. These include both primary tumors of the sacrum and locally invasive colorectal carcinomas. Sacral resection is often the only effective alternative for meaningful palliation or cure of sacral tumors. A review of 20 cases of sacral resections for primary sacral tumors (8) and locally invasive anorectal cancers (12) is presented. The mortality (0%) and morbidity (35% urinary complications, 25% wound disruptions, 1600-mL median blood loss) compare favorably with reports in the literature. Long-term survival was achieved with primary tumors of the sacrum. Local control of disease was achieved in the majority of patients with rectal cancer, with good palliation of preoperative pain. Long-term survival, however, is rare in this group. Surgical resection of sacral tumors can be undertaken with acceptable morbidity and mortality in selected patients.


Suboptimal Dosing of Fluconazole in Critically Ill Patients: Time To Rethink Dosing.
Fluconazole is frequently used for the treatment of invasive Candida infections in critically ill patients. However, alterations in renal functions might influence fluconazole clearance. Therefore, our objective was to study the impact of renal function on the population pharmacokinetics of fluconazole in critically ill patients with various degrees of renal function or undergoing continuous renal replacement therapy (CRRT). This was an open-label, multicenter observational study. Critically ill patients receiving fluconazole were included. Baseline and clinical data were collected. At days 3 and 7 of enrollment, blood samples were drawn for pharmacokinetic curves. Additionally, daily trough samples were taken. A nonlinear mixed-effects model was built, followed by Monte Carlo simulations for assessment of exposure to various dosages of fluconazole. Nineteen patients were included with a median age of 64.4 (range, 23 to 81) years and median weight of 82.0 (range, 44.0 to 119.5) kg. A linear two-compartment model best described fluconazole pharmacokinetics and demonstrated higher clearance than expected in critically ill patients. Simulations showed that daily dosages of 600 mg and 800 mg are needed for intensive care unit (ICU) patients with normal renal function and patients on CRRT, respectively, to achieve the EUCAST-recommended target fAUC (area under the concentration-time curve for the free, unbound fraction of the drug)/MIC ratio of 100. In conclusion, fluconazole clearance is highly variable in ICU patients and is strongly dependent on renal function and CRRT. Trough concentrations correlated well with the AUC, opening up opportunities for tailored dosing using therapeutic drug monitoring. We recommend doses of 400 mg for patients with poor to moderate renal function, 600 mg for patients with adequate renal function, and 800 mg for patients treated with CRRT. (This study has been registered at ClinicalTrials.gov under identifier NCT02666716.).


Immunoassay of haemoglobin-acrylonitrile adduct in rat as a biomarker of exposure.
Acrylonitrile (AN) is a rat carcinogen. Human exposure may come from chemical industries and smoking. A haemoglobin adduct of acrylonitrile (Hb-AN) has been used as a biomarker of exposure by means of gas chromatography-mass spectrometry (GC-MS) analysis. We have developed specific monoclonal antibodies (Mab) to human Hb-AN and wish to report evaluation of an immunoassay in rats using an Mab that cross-reacts with rat Hb-AN. A dose response study of LD 0, 10, 50, and 90 in Sprague-Dawley rats was undertaken, with each rat receiving \[2,3-14C]AN at 50 Ci kg-1 sc, and Hb from an aliquot of blood was taken for covalent binding analysis by liquid scintillation spectrometry and fluorescence ELISA. The dose responses of rats at 0.25, 0.5, 1.0, and 2.0 h after AN doses of 20, 50, 80, 115 mg kg-1 were compared by both methods with Hb and globin samples. Regression analysis showed a linear relationship between immunoassay and 14C-AN binding. This indicates that an antigenic form of Hb-AN may be used as a surrogate of Hb-AN adduct. The sensitivity of ELISA was tested in rats exposed for 1 h to sub-toxic doses of AN (10-1.1 mg kg-1). Quantification of Hb-AN by immunoassay was achieved by calibration with a synthetic adduct HbAN4h, a reference adduct prepared by treating rat Hb with excess AN for 4 h. ELISA and GC-MS analysis of N-terminal valine-AN in the Hb-AN adduct were compared and similar detection levels were found. This rat study appears to have validated the new immunoassay method for biomonitoring of AN exposure.


Role of adenosine and N-methyl-D-aspartate receptors in mediating haloperidol-induced gene expression and catalepsy.
Acute blockade of dopamine D(2) receptors by the typical antipsychotic drug haloperidol leads to alterations in neuronal gene expression and behavior. In the dorsolateral striatum, the levels of mRNA for the immediate-early gene c-fos and the neuropeptide gene neurotensin/neuromedin N (NT/N) are significantly increased by haloperidol. An acute behavioral response to haloperidol is catalepsy, considered to be a rodent correlate of some of the immediate extrapyramidal motor side effects seen in humans. Several lines of evidence suggest a link between neurotensin induction in the dorsolateral striatum and catalepsy. We hypothesize that both striatal gene induction and catalepsy elicited by haloperidol arise from the combined effect of excitatory adenosinergic and glutamatergic inputs acting at adenosine A(2A) and N-methyl-D-aspartate (NMDA) receptors, respectively. In agreement with our previous reports, adenosine antagonists reduced haloperidol-induced c-fos and neurotensin gene expression as well as catalepsy. In agreement with other reports, the noncompetitive NMDA receptor antagonist MK-801 also reduced gene expression and catalepsy in response to haloperidol. The competitive NMDA receptor antagonist LY235959 decreased haloperidol-induced catalepsy. We show here that blocking both A(2A) and NMDA receptors simultaneously in conjunction with haloperidol resulted in a combined effect on gene expression and behavior that was greater than that for block of either receptor alone. Both c-fos and NT/N mRNA levels were reduced, and catalepsy was completely abolished. These results indicate that the haloperidol-induced increases in c-fos and NT gene expression in the dorsolateral striatum and catalepsy are driven largely by adenosine and glutamatergic inputs acting at A(2A) and NMDA receptors.


A modified tensionless gingival grafting technique using acellular dermal matrix.
Conventional surgical procedures designed for autogenous tissue material may not be appropriate when using acellular dermal matrix (ADM) for the treatment of gingival recessions. This article describes a new surgical technique that addresses the unique and sensitive aspects of ADM specifically to improve esthetic outcomes and gain increased clinical predictability when treating Miller Class I and II gingival recession defects. In this paper, a root coverage case is described and the specific steps and rationale for this new technique are explained. This technique has been predictable clinically, with results comparable to those achieved using autogenous tissue.


[A case of primary seminal vesicle cancer detected by FDG-PET/CT].
We reported a case of primary seminal vesicle cancer, detected by FDG-PET/CT. A 65-year-old man with constipation and appetite loss was admitted to our hospital. An ultrasound examination revealed evidence of bilateral hydronephrosis. He was diagnosed as acute post renal failure, and nephrostomy was done. CT and MRI showed a solid mass in the area of seminal vesicle. He underwent transrectal core biopsy, which histologically showed poorly differentiated adenocarcinoma. Immunohistochemistry showed the tumor to be CA125 positive, CEA positive and CK7 positive but PSA negative. FDG-PET/CT revealed an increased uptake of FDG only in the area of seminal vesicle. Serum CA125 was elevated and PSA stayed within normal limit. Primaly rectal carcinoma was ruled out by colonoscopy. The result of transperineal prostate biopsy was negative. We diagnosed him as suffering from primary seminal vesicle carcinoma. Anti-androgen blockade and radiotherapy to whole pelvis were performed, and serum CA125 level was improved. But, 6 months later serum CA125 re-elevated and 19 months later multiple liver metastases were noted. The patient received two cycles of docetaxel and cisplatin chemotherapy, however he developed pulmonaly embolism and rectal bleeding by tumor invasion and he died of his disease 22 months after the diagnosis.


The relationship between lumbar spine load and muscle activity during extensor exercises.
There have been no previous studies that quantitatively assessed the load on the spine during extensor exercises. The purpose of our study was to investigate the loading of the lumbar spine and trunk muscle activity levels while subjects performed typical trunk extensor exercises. Thirteen male volunteers (mean age = 21.0 years, SD = 1.0, range = 19-23; mean height = 176.0 cm, SD = 6.2, range = 165-188; mean mass = 77.0 kg, SD = 7.0, range = 63-89) participated. The subjects performed four different back exercises. Electromyographic (EMG) activity was recorded from 14 trunk muscles. The postures that corresponded to the maximum external moment were identified and quantified using rigid body modeling combined with an EMG-driven model to determine joint loading at the L4-5 joint. The exercises were then evaluated based on the lumbar spine loading and peak muscle activity levels. A reference task of lifting 10 kg from midthigh was included for comparison. The exercises involving active trunk extension produced the highest joint forces and muscle activity levels. Exercises involving leg extension with the spine held isometrically demonstrated asymmetrical activity of the trunk muscles, thereby reducing loads on the spine. The back extensor exercises examined provided a wide range of joint loading and muscle activity levels. Single-leg extension tasks appear to constitute a low-risk exercise for initial extensor strengthening, given the low spine load and mild extensor muscle challenge. When combined with contralateral arm extensions, the challenge and demand of the exercise were increased. The compressive loading and extensor muscle activity levels were highest for the trunk extension exercises.


Hollow Gaussian beams and their propagation properties.
A new mathematical model, described as hollow Gaussian beams (HGBs), is proposed to describe a dark hollow laser beam (DHB). The area of the dark region across the HGBs can easily be controlled by proper choice of the beam parameters. Based on the Collins integral, an analytical propagation formula for the HGBs through a paraxial optical system is derived. The HGBs also can be expressed as a superposition of a series of Lagurerre-Gaussian modes by use of a polynomial expansion. As a numerical example, the propagation properties of a DHB in free space are illustrated graphically. The HGBs provide a convenient and powerful way to describe and treat the propagation of DHBs and can be used conveniently to analyze atoms manipulated with a DHB.


In vitro evaluation of tropolone absorption, metabolism, and clearance.
Tropolone compounds can inhibit hepatitis B virus (HBV) replication at sub-micromolar levels and are synergistic upon co-treatment with nucleos(t)ide analog drugs. However, only a few compounds within this chemotype have been screened for their pharmacological properties. Here, we chose 36 structurally diverse tropolones from six subclasses to characterize their in vitro pharmacological parameters. All compounds were more soluble in pHs that reflect the gastrointestinal tract (pH 5 and 6.5) than plasma (pH 7.4). Those compounds that had solubility limits >100 μM were tested in a passive permeability assay, and there was no general trend in the compounds' passive permeability at any pH. Twenty-nine compounds with the best absorption parameters were tested in HEK293 cells to assess potential cytotoxicity; measured toxicities were similar to those in the hepatic HepDES19 cells used for screening (R2 = 0.55). Sixteen representative compounds were tested against five major CYP450 isoforms and there was no substantial inhibition by any compound against any of the enzymes tested (<50%). The t1/2 values of 15 compounds were determined in the microsome stability assay and 12 compounds were evaluated in plasma protein binding assays to assess factors affecting their rate of clearance. All compounds with detectable analyte peaks had t1/2 > 30 min, and while 4 of 12 had statistically significant decreased potency in conditions with increased albumin concentrations, only one compound's potency was biologically significant. These data indicate that the tropolones have pharmacological characteristics that reflect approved drugs and inform future structure activity relationships during drug design.


Cardiac and kidney markers for cardiovascular prediction in individuals with chronic kidney disease: the Atherosclerosis Risk in Communities study.
Traditional predictors suboptimally predict cardiovascular disease (CVD) in individuals with chronic kidney disease (CKD). This study compared 5 nontraditional cardiac and kidney markers on the improvement of cardiovascular prediction among those with CKD. Among 8622 participants aged 52 to 75 years in the Atherosclerosis Risk in Communities (ARIC) Study, cardiac troponin T, N-terminal pro-B-type natriuretic peptide, cystatin C, β2-microglobulin, and β-trace protein were compared for improvement in predicting incident CVD after stratifying by CKD status (940 participants with CKD [kidney dysfunction or albuminuria]). During a median follow-up of 11.9 years, there were 1672 CVD events including coronary disease, stroke, and heart failure (336 cases in CKD). Every marker was independently associated with incident CVD in participants with and without CKD. The adjusted hazard ratios (per 1 SD) were larger for cardiac markers than for kidney markers, particularly in CKD (1.61 [95% confidence interval, 1.43-1.81] for cardiac troponin T, 1.50 [1.34-1.68] for N-terminal pro-B-type natriuretic peptide, and <1.26 for kidney markers). Particularly in CKD group, cardiac markers compared with kidney markers contributed to greater c-statistic increment (0.032-0.036 versus 0.012-0.015 from 0.679 with only conventional predictors in CKD and 0.008-0.011 versus 0.002-0.010 from 0.697 in non-CKD) and categorical net reclassification improvement (0.086-0.127 versus 0.020-0.066 in CKD and 0.057-0.077 versus 0.014-0.048 in non-CKD). The superiority of cardiac markers was largely consistent in individual CVD outcomes. A greater improvement in cardiovascular prediction was observed for cardiac markers than for kidney markers in people with CKD. These results suggest that cardiac troponin T and N-terminal pro-B-type natriuretic peptide are useful for better CVD risk classification in this population.


[A rare presentation of tuberculosis. Miliary tuberculosis in a immunocompetent patient].
Currently, miliary TB has a higher incidence among the elderly and immunodepressed patients, and exceptionally in young adults. A case of miliary tuberculosis was diagnosed in a young immunocompetent patient without using invasive techniques. The clinical and diagnostic implications will be discussed.


Intermolecular interactions and morphology of aqueous polymer/surfactant mixtures containing cationic chitosan and nonionic sorbitan esters.
In this study, the impact of surfactant molecular composition (saturated sorbitan monolaurate or unsaturated sorbitan monooleate) on polymer/surfactant assemblies was examined. Specifically, the associations between the cationic chitosan and the uncharged surfactants were monitored by surface tension, turbidity, and conductivity measurements. Bright field, confocal laser scanning, and transmission electron microscopy revealed that nanometer-sized chitosan/surfactant aggregates comprised of a chitosan-rich shell and a chitosan-poor core agglomerate at high surfactant concentrations to yield micrometer-scaled supramolecular structures with highly ordered internal structure. The size and architecture of these chitosan/surfactant assemblies were dependent on the structure and concentration of the surfactant employed. The association mechanism among chitosan, surfactant, and the chitosan/surfactant aggregates was discussed in terms of the semirigid polyelectrolyte character of chitosan and the hydrophobic character of sorbitan esters. This study provides important insight into the structural and physical parameters of surfactant that govern the formation of multicompartment polymer/surfactant assemblies.


Targeted inhibition of the deubiquitinating enzymes, USP14 and UCHL5, induces proteotoxic stress and apoptosis in Waldenström macroglobulinaemia tumour cells.
Deubiquitinase enzymes (DUBs) of the proteasomal 19S regulatory particle are emerging as important therapeutic targets in several malignancies. Here we demonstrate that inhibition of two proteasome-associated DUBs (USP14 and UCHL5) with the small molecule DUB inhibitor b-AP15, results in apoptosis of human Waldenström macroglobulinaemia (WM) cell lines and primary patient-derived WM tumour cells. Importantly, b-AP15 produced proteotoxic stress and apoptosis in WM cells that have acquired resistance to the proteasome inhibitor bortezomib. In silico modelling identified protein residues that were critical for the binding of b-AP15 with USP14 or UCHL5 and proteasome enzyme activity assays confirmed that b-AP15 does not affect the proteolytic capabilities of the 20S proteasome β-subunits. In vitro toxicity from b-AP15 appeared to result from a build-up of ubiquitinated proteins and activation of the endoplasmic reticulum stress response in WM cells, an effect that also disrupted the mitochondria. Focused transcriptome profiling of b-AP15-treated WM cells revealed modulation of several genes regulating cell stress and NF-κB signalling, the latter whose protein translocation and downstream target activation was reduced by b-AP15 in vitro. This is the first report to define the effects and underlying mechanisms associated with inhibition of USP14 and UCHL5 DUB activity in WM tumour cells.


Cognitive behavioural interventions for sleep problems in adults aged 60+.
The prevalence of sleep problems in adulthood increases with age. While not all sleep changes are pathological in later life, severe disturbances may lead to depression, cognitive impairments, deterioration of quality of life, significant stresses for carers and increased healthcare costs. The most common treatment for sleep disorders (particularly insomnia) is pharmacological. The efficacy of non-drug interventions has been suggested to be slower than pharmacological methods, but with no risk of drug-related tolerance or dependency. Cognitive and behavioural treatments for sleep problems aim to improve sleep by changing poor sleep habits, promoting better sleep hygiene practices and by challenging negative thoughts, attitudes and beliefs about sleep. To assess the efficacy of cognitive-behavioural interventions in improving sleep quality, duration and efficiency amongst older adults (aged 60 and above). The following databases were searched: MEDLINE (1966 - October 2001); EMBASE (1980 - January 2002), CINAHL ( 1982 - January 2002; PsychINFO 1887 to 2002; The Cochrane Library (Issue 1, 2002); National Research Register (NRR [2002]). Bibliographies of existing reviews in the area, as well as of all trial reports obtained, were searched. Experts in the field were consulted. Randomised controlled trials of cognitive behavioural treatments for primary insomnia where 80% or more of participants were over 60. Participants must have been screened to exclude those with dementia and/or depression. Abstracts of studies identified in searches of electronic databases were read and assessed to determine whether they might meet the inclusion criteria. Data were analysed separately depending on whether results had been obtained subjectively or objectively. Six trials, including 282 participants with insomnia, examined the effectiveness of cognitive-behavioural treatments (CBT) for sleep problems in this population. The final total of participants included in the meta-analysis was 224. The data suggest a mild effect of CBT for sleep problems in older adults, best demonstrated for sleep maintenance insomnia. When the possible side-effects of standard treatment (hypnotics) are considered, there is an argument to be made for clinical use of cognitive-behavioural treatments. Research is needed to establish the likely predictors of success with such treatments. As it may well be the case that the treatment efficacy of cognitive-behavioural therapy itself is not durable, the provision of "top-up" sessions of CBT training to improve durability of effect are worthy of investigation.


Enduring effects of evidence-based psychotherapies in acute depression and anxiety disorders versus treatment as usual at follow-up--a longitudinal meta-analysis.
This meta-analysis examined the enduring efficacy of evidence-based psychotherapies (EBP) in comparison to treatment as usual (TAU) by examining effects from termination to follow-up for acute anxiety and depression in an adult outpatient population. It was hypothesized that EBPs might extend their efficacy at follow-up assessment (Tolin, 2010). Longitudinal multilevel meta-analyses were conducted that examined the magnitude of difference between EBP and TAU. Targeted (disorder-specific) outcomes were examined, along with dropout rates at follow-up assessments. A total of 15 comparisons (including 30 repeated effect sizes [ES]) were included in this meta-analysis (average of 8.9 month follow-up). Small to moderate ES differences were found to be in favor of EBPs at 0-4 month assessments (Hedges' g=0.40) and up to 12-18 month assessments (g=0.20), indicating no extended efficacy at follow-up. However, the TAU-conditions were heterogeneous, ranging from absence of minimal mental health treatment to legitimate psychotherapeutic interventions provided by trained professionals, the latter of which resulted in smaller ES differences. Furthermore, samples where substance use comorbidities were not actively excluded indicated smaller ES differences. TAU-conditions produced slightly higher dropout rates than EBP-conditions. Findings indicate small and no extended superiority of EBP for acute depression and anxiety disorders in comparison to TAU at follow-up assessment. There are a limited number of studies investigating the transportability and lasting efficacy of EBP compared to TAU, especially to TAU with equivalent conditions between treatment groups.


[Tumor characteristics and five-year survival in breast cancer patients at the National Cancer Institute, Rio de Janeiro, Brazil].
Numerous factors have been studied to establish more secure prognostic criteria in breast cancer patients. This study estimates five-year survival rates and principal prognostic factors related to tumor characteristics in women with invasive breast cancer and submitted to surgery at the National Cancer Institute, Rio de Janeiro, Brazil, from May 1995 to July 1996. Study variables were: lymph node status, tumor size, aggressiveness grade, and presence of estrogen and progesterone receptors. Survival functions were calculated according to the Kaplan-Meyer method. The Cox proportional hazards model was used to evaluate prognostic factors. Five-year survival was 75% for all women and 64% for those with node involvement. Multivariate analyses identified node involvement as the strongest predictor of outcome; a positive estrogen receptor test was associated with a better prognosis. These findings highlight the need for studies to assess new variables to be added to known factors in order to better orient therapy for breast cancer.


[Treatment of post-herpes zoster pain with tramadol. Results of an open pilot study versus clomipramine with or without levomepromazine].
To date, no universally applicable recommendations are available for the treatment of patients with postherpetic neuralgia. A mixture of clinical anecdotes, experimental findings and observations from clinical trials form the basis of the medical arsenal for this condition. Tricyclic antidepressants are commonly used, and clinical experience and several investigations have documented their effectiveness. Today, single entity antidepressants, which can be combined with neuroleptics to increase analgesia, are generally recommended for the treatment of postherpetic neuralgia. Some authors also recommend the additional administration of an opioid if analgesia is inadequate. Just over a decade ago, opioids were considered ineffective for the treatment of neuropathic pain; however, more recent investigations relating to the use of opioids, primarily in the treatment of nontumour-related chronic pain, have led to a revision of their use in neuropathic pain. Nevertheless, the use of opioid therapy for neurogenic pain remains controversial. Tramadol is a synthetic, centrally acting analgesic with both opioid and nonopioid analgesic activity. The nonopioid component is related to the inhibition of noradrenaline (norepinephrine) reuptake and stimulation of serotonin (5-hydroxytryptamine; 5-HT) release at the spinal level. In this regard, there are parallels with antidepressants, which are believed to potentiate the effect of biogenic amines in endogenous pain-relieving systems. There is evidence that, in tramadol, both mechanisms act synergistically with respect to analgesia. The aim of this pilot study was to investigate, for the first time, the analgesic efficacy and tolerability of tramadol, compared with the antidepressant clomipramine, in the treatment of postherpetic neuralgia. If necessary, clomipramine was used in combination with the neuroleptic levomepromazine. The study allowed individualised dosages at predetermined intervals up to a maximum daily dose of tramadol 600mg and clomipramine 100mg, or clomipramine 100mg with or without levomepromazine 100mg. 21 (60%) of 35 randomised patients (> or = 65 years) received the study medication over the 6-week period [tramadol n = 10; clomipramine with or without levomepromazine) n = 11]. After 3 weeks' treatment the dosage in both groups remained almost constant for the rest of the 6-week treatment phase (mean daily dose: tramadol 250 to 290mg; clomipramine 59.1 to 63.6mg). Only 3 patients required the combination of clomipramine and levomepromazine. At the outset, both groups recorded an average pain level of 'moderate' to 'very severe'. In correlation with increasing the study medication, this had decreased to 'slight' by the end of the treatment, when 9 of 10 patients in the tramadol group and of 6 of 11 patients in the clomipramine group retrospectively rated their analgesia as excellent, good or satisfactory. The psychological/physical condition of the patients did not change significantly during tramadol treatment. Sensitivity and depression parameters decreased in the clomipramine group. The incidence of adverse events for all patients was similar in both groups (tramadol 76.5%; clomipramine with or without levomepromazine 83.3%). In conclusion, tramadol would appear to be an interesting therapeutic alternative for pain relief in postherpetic neuralgia, particularly in patients who are not depressed. In clinical practice, tramadol and clomipramine can best be used differentially. For example, tramadol could be the drug of first choice in patients with obvious cardiovascular disease (not an uncommon problem in the > or = 65 year age group) in whom antidepressants are contraindicated, and similarly in patients in whom an antidepressant effect is not required. (ABSTRACT TRUNCATED)


Identification of a common enterobacterial flagellin epitope with a monoclonal antibody.
A monoclonal antibody (mAb), designated 15D8, was produced from BALB/c splenocytes of mice injected with Escherichia coli flagella. ELISA of motile cells, non-motile cells and partially purified flagellin proteins showed that the mAb reacted specifically with flagella of E. coli and with other members of the family Enterobacteriaceae. Western immunoblot analyses of enterobacterial flagella or cell extracts demonstrated that the antibody reacted with a single protein species in the extracts which was identical in size to purified flagellin. The antigenic determinant for this antibody appears to be surface exposed and linear in configuration, since the antibody reacted with native flagella and flagella which had been denatured. This antibody was also used to demonstrate that although the flagella proteins are heterogeneous in size, at least one epitope is highly conserved.


Are Upper-Body Axial Symptoms a Feature of Early Parkinson's Disease?
Axial disorders are considered to appear late in the course of Parkinson's disease (PD). The associated impact on quality of life (QoL) and survival and the lack of an effective treatment mean that understanding and treating axial disorders is a key challenge. However, upper-body axial disorders (namely dysarthria, swallowing and breathing disorders) have never been prospectively assessed in early-stage PD patients. To characterize upper-body axial symptoms and QoL in consecutive patients with early-stage PD. We prospectively enrolled 66 consecutive patients with early-stage PD (less than 3 years of disease progression) and assessed dysarthria, dysphagia and respiratory function (relative to 36 controls) using both objective and patient-reported outcomes. The mean disease duration was 1.26 years and the mean UPDRS motor score was 19.4 out of 108. 74% of the patients presented slight dysarthria (primarily dysprosodia). Men appeared to be more severely affected (i.e. dysphonia). This dysfunction was strongly correlated with low swallowing speed (despite the absence of complaints about dysphagia), respiratory insufficiency and poor QoL. Videofluorography showed that oral-phase swallowing disorders affected 60% of the 31 tested patients and that pharyngeal-phase disorders affected 21%. 24% of the patients reported occasional dyspnea, which was correlated with anxiety in women but not in men. Marked diaphragmatic dysfunction was suspected in 42% of the patients (predominantly in men). Upper body axial symptoms were frequent in men with early-stage PD, whereas women presented worst non-motor impairments. New assessment methods are required because currently available tools do not reliably detect these upper-body axial disorders.


Correction of congenital hydrocephalus in utero II: Efficacy of in utero shunting.
To study the effect of in utero ventricular decompression of hydrocephalus on brain development and prognosis, and to evaluate the function and possible complications of different shunt designs, we created fetal hydrocephalus in 28 fetal lambs and 17 fetal monkeys by injecting kaolin into the cisterna magna during the third trimester. One fetal lamb had indwelling intracranial subdural and amniotic cavity pressure catheters placed to study serial changes in the relationship of intracranial pressure (ICP) and amniotic fluid pressure (AFP) from the time of injection until term. Twenty hydrocephalic fetal lambs underwent ventricular decompression--ventriculoamniotic (V-A), N = 10; ventriculo-right atrial (V-RA), N = 9; and ventriculo-pleural (V-PL), N = 1--21 to 25 days after the kaolin was injected; seven hydrocephalic fetuses were left unshunted as controls (CON). Eight fetal monkeys underwent V-A decompression 14 to 21 days after kaolin injection; nine were left unshunted as controls. All animals were delivered by cesarean section near term, assessed for viability and their brains examined grossly and microscopically. After kaolin injection in the fetal lamb, ICP (r = 0.94) and ICP minus AFP (r = 0.93) rose in a linear fashion, while AFP showed no trend (r = 0.22). All unshunted newborn lambs and monkeys had split sutures, dilated ventricles, and thinned cortical mantle. Most shunted lambs showed anatomic improvement with decreased head circumference, overriding sutures, normal-sized ventricles, and improved survival. However, histopathology revealed marked white matter destruction. In contrast, most shunted monkeys showed little anatomic improvement. Both shunted and unshunted monkey brains showed a severe inflammatory ventriculitis. In both lambs and monkeys, shunting was associated with a variety of complications including subdural hematoma, subdural hygroma, shunt infection, shunt occlusion, and improper shunt tip placement. In this experimental model, in utero decompression of obstructive hydrocephalus improves overall survival, improves gross ventriculomegaly, does not improve histopathologic brain damage, and is associated with significant complications. The choice of the optimal shunt design and the effect on postnatal neurologic function requires further study. This work emphasizes the need for continuing research in an animal model prior to human application.


Synthesis and characterization of cyclohexane-1R,2R-diamine-based Pt(iv) dicarboxylato anticancer prodrugs: their selective activity against human colon cancer cell lines.
Three pairs of asymmetric dicarboxylato derivatives based on the cisplatin and oxaliplatin-like skeletons have been synthesized de novo or re-synthesized. The axial ligands consist of one medium-chain fatty acid (MCFA), namely clofibrate (i.e. 2-(p-chlorophenoxy)-2-methylpropionic acid, CA), heptanoate (HA) or octanoate (OA), respectively, and an inactive acetato ligand that imparts acceptable water solubility to such conjugates. Stability tests provided evidence for the partial formation of two hydrolyzed products, corresponding to two monoaqua diastereomers derived from the substitution of an equatorial chlorido ligand with a water molecule. The complexes have been tested on three different colon cancer cell lines having different histological history, and also on the cisplatin-sensitive A2780 ovarian cancer cell line for comparison. This allowed the evaluation not only of the increase in activity on passing from Pt(ii) to Pt(iv) derivatives, but also the selectivity towards colon cancer cells brought about by the cyclohexane-1R,2R-diamine carrier ligand.