Bilateral femoral neuropathy after vaginal delivery: A case report.
Epidural analgesia is frequently used for the management of labor pain in parturients. The neurologic complications of epidural analgesia are rare, but they are catastrophic when they occur. We report here on a case of bilateral femoral neuropathy in a vaginal delivery patient with epidural analgesia. The patient complained of weakness of both thighs and numbness around both knees. Neurologic examination and electromyographic study revealed that they occurred due to the delivery itself. The patient had not fully recovered at eight weeks after delivery. The declining incidence of postpartum femoral neuropathy may reflect a reduced duration of labor in accordance with modern obstetric practice, and particularly the more frequent use of Caesarean delivery. The exact etiology of postpartum femoral neuropathy is unknown, and the possible factors are direct compression of nerves that are proximal within the pelvis by either the fetal head or birthing instruments, or this malady is the result of pressure induced ischemia at the level of the inguinal ligament when the pregnant woman is in the lithotomy position.


Structural characterization of the K antigens from Rhizobium fredii USDA257: evidence for a common structural motif, with strain-specific variation, in the capsular polysaccharides of Rhizobium spp.
Rhizobium fredii participates in a nitrogen-fixing symbiosis with soybeans, in a strain-cultivar-specific interaction, and past studies have shown that the cell surface and extracellular polysaccharides of rhizobia function in the infection process that leads to symbiosis. The structural analysis of the capsular polysaccharides (K antigens) from strain USDA257 was performed in this study. The K antigens were extracted from cultured cells with hot phenol-water and purified by size exclusion chromatography. We isolated two structurally distinct K antigens, both containing a high proportion of 3-deoxy-D-manno-2-octulosonic acid (Kdo). The polysaccharides were characterized by matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry, nuclear magnetic resonance spectrometry, and gas chromatography-mass spectrometry analyses. The primary polysaccharide, which constituted about 60% of the K-antigen preparation, consisted of repeating units of mannose (Man) and Kdo, [-->)3-beta-D-Manp-(1-->5)-beta-D-Kdop-(2-->], and a second polysaccharide consisted of 2-O-MeMan and Kdo, [-->)3-beta-D-2-O-MeManp-(1-->5)-beta-D-Kdop-(2-->]. These structures are similar to yet distinct from those of other strains of R. fredii and R. meliloti, and this finding provides further evidence that the K antigens of rhizobia are strain-specific antigens which are produced within a conserved motif.


Synergistic effect of cryptotanshinone and temozolomide treatment against human glioblastoma cells.
Glioblastoma multiforme (GBM) is a complex disease to treat owing to its profound chemoresistance. Therefore, we evaluated the combined effect and therapeutic efficacy of temozolomide (TMZ), a potent alkylating agent and the current gold standard therapy for GBM, and cryptotanshinone (CTS), which inhibits glioma cell proliferation in GBM cells. Using LN229 and U87-MG human GBM cells in a short-term stimulation in vitro model, the cytotoxic and anti-proliferative effects of single and combined treatment with 4 μM CTS and 200 μM TMZ were investigated. Furthermore, cell viability, DNA damage, apoptosis rate, and signal transducer and activator of transcription 3 (STAT3) protein were measured using cytotoxic assay, comet assay, flow cytometry, and western blotting analysis, respectively. The two drugs' synergistic interaction was validated using the synergy score. We found that the anti-proliferative effects of combination therapy using the two drugs were greater than that of each agent used alone (CTS or TMZ). Western blot analysis indicated that treatment of GBM cells with CTS combined with TMZ more significantly decreased the expression of MGMT and STAT3, than that with TMZ alone. Combined treatment with CTS and TMZ might be an effective option to overcome the chemoresistance of GBM cells in a long-term treatment strategy.


Bird sky networks: how do avian scavengers use social information to find carrion?
The relative contribution of personal and social information to explain individual and collective behavior in different species and contexts is an open question in animal ecology. In particular, there is a major lack of studies combining theoretical and empirical approaches to test the relative relevance of different hypothesized individual behaviors to predict empirical collective patterns. We used an individual-based model to confront three hypotheses about the information transfer between social scavengers (Griffon Vultures, Gyps fulvus) when searching for carrion: (1) Vultures only use personal information during foraging ("nonsocial" hypothesis); (2) they create long chains of vultures by following both other vultures that are flying towards carcasses and vultures that are following other vultures that are flying towards carcasses ("chains of vultures" hypothesis); and (3) vultures are only attracted by other vultures that are sinking vertically to a carcass ("local enhancement" hypothesis). The chains of vultures hypothesis has been used in existing models, but never been confronted with field data. Testing is important, though, because these hypotheses could have different management implications. The model was parameterized to mimic the behavior and the densities of both Griffon Vultures and carcasses in a 10,000-km2 study area in northeastern Spain. We compared the number of vultures attending simulated carcasses with those attending 25 continuously monitored experimental carcasses in the field. Social hypotheses outperformed the nonsocial hypothesis. The chains of vultures hypothesis overestimated the number of vultures feeding on carcasses; the local enhancement hypothesis fitted closely to the empirical data. Supported by our results, we discuss mechanistic and adaptive considerations that reveal that local enhancement may be the key social mechanism behind collective foraging in this and likely other avian scavengers and/or social birds. It also highlights the current need for more studies confronting alternative models of key behaviors with empirical patterns in order to understand how collective behavior emerges in animal societies.


Long-term follow-up of laparoscopic repair of parastomal hernia using a bilayer mesh with a slit.
Open surgery for parastomal hernia has been associated with high morbidity and recurrence rates exceeding 50%. Laparoscopic mesh repair is a promising alternative. Published series on laparoscopic mesh repair of parastomal hernia, however, are few with relative short follow-up. Seventy-two consecutive patients with 48 paracolostomy and 24 paraileostomy hernias were studied prospectively. Using a two-layer mesh with polypropylene on the parietal side and nonadhesive PTFE toward the viscera, a slit with a central keyhole was cut in the mesh modified after Hofstetter. Covering the fascial defect, the slit was closed laterally. Of the 70 patients discharged alive, 66 were followed for 6 months to 11 years (median=3 years). Four patients were lost for follow-up. Lysis of adhesions to the anterior abdominal wall, necessary in 68 patients (94%), was frequently a cumbersome and time-consuming task. In nine patients it was the apparent cause of inadvertent full-thickness enterotomy and seromuscular lesion in three and six patients, respectively. Three procedures (4%) were converted to open surgery. Postoperative complications were observed in 16 patients (22%), nine of whom required repeat surgery (13%). Two patients (3%) died. The median hospital stay was 3 days. Late mesh-related complications were observed in five patients (7%) after 5-34 months. Three patients developed abscess that required mesh removal in two patients, including one patient with a small bowel fistula. In two patients the mesh was removed because of small-bowel obstruction and stenosis of the colon at the level of the mesh, respectively. Parastomal hernia recurred in two patients 1 month and 52 months after surgery (recurrence rate=3%, 95% confidence interval=1-10). Laparoscopic repair of parastomal hernia using bilayer mesh with a slit is associated with a risk of substantial postoperative morbidity, including late mesh-related complications, but a recurrence rate of less than 10%.


Flipping up the board: Losing my mother to suicide.
Personal account of losing a parent due to a firearm and a completed suicide attempt related to mental health crisis.


Draft Genome Sequences of Idiomarina abyssalis Strain KJE, Marinobacter salarius Strain NP2017, and Marinobacter salarius Strain AT3901, Isolated from Deep-Sea Sediment near the Western Flank of the Mid-Atlantic Ridge.
We report the draft genomes of environmental cultures collected from shallow sediment from the western flank of the Mid-Atlantic Ridge. The isolates were most closely related to Idiomarina abyssalis strain KJE (100% complete), Marinobacter salarius strain NP2017 (97.6% complete), and Marinobacter salarius strain AT3901 (98.4% complete). Isolates identified as an Idiomarina species possess complete nitrite oxidation and reduction pathways, and isolates identified as a Marinobacter species possess complete dissimilatory nitrate reduction pathways.


Long-Term Follow-Up of a Case with Dyskeratosis Congenita Caused by NHP2-V126M/X154R Mutation: Genotype-Phenotype Association.
Dyskeratosis congenita (DC) is a rare inherited syndrome characterized by classical mucocutaneous features and the presence of other clinical features including bone marrow failure, pulmonary fibrosis, liver cirrhosis, and a predisposition to cancer. The symptoms develop at various ages and may manifest over time. Gene mutations associated with DC, such as DC1, TERC, TERT, TINF2, NHP2, NOP10, ACD, CTC1, NAF1, PARN, POT1, RTEL1, STN1, and WRAP53, have been identified in about 70% of patients. Since the number of patients with DC is small and the effect of genetic pathogenic variant may affect the phenotype, we wanted to present the clinical features and course of illness in a patient with NHP2 gene mutation (compound heterozygote for the NHP2 mutations c.376G>A/c.460T>A; amino acid substitutions: p.Val126Met and p.X154Arg) that occurred as a compound heterozygous state.


Secondhand Smoke and Streptococcal Infection in Young Children Under Japan's Voluntary Tobacco-Free Policy.
Tobacco-free policy in Japan lags behind those of most developed countries. Evidence is required to promote strong implementation of existing policies. This study aimed to assess whether exposure to secondhand smoke (SHS) influences the incidence of streptococcal infection in young children, to further support the need for effective tobacco-free policies in Japan. This study used medical administrative claim and health check data from the Japan Health Insurance Association Fukuoka branch. Participants were beneficiaries' dependents younger than age 4 years. Exposure was defined as SHS from beneficiaries' smoking, each year during 2011-2014. The outcome was incidence of streptococcal infection, diagnosed with and without laboratory testing. Logistic regression analysis was performed to yield odds ratios (ORs) of associations with the outcome and 95% confidence intervals (CIs). This study included a total of 5743 children. The proportion of all participants with a record of streptococcal infection was 4.2% (n = 244). The results of logistic regression analysis between streptococcal infection and SHS exposure showed a significantly higher association (OR 1.39, 95% CI 1.07-1.80, P < 0.05) if all cases were included and an insignificant association with diagnoses using testing (OR 1.20, 95% Cl 0.80-1.80, P = 0.39). This study showed that 60% of streptococcal infections in young children were diagnosed without testing, and SHS increased this incidence regardless of testing. It reports new findings regarding the effect of SHS on infection in young children to support implementation and promotion of tobacco-free policies by the Japanese government not only in public spaces, but also at home.


Bardoxolone methyl analog attenuates proteinuria-induced tubular damage by modulating mitochondrial function.
Multiple clinical studies have shown that bardoxolone methyl, a potent activator of nuclear factor erythroid 2-related factor 2 (Nrf2), is effective in increasing glomerular filtration rate in patients with chronic kidney disease. However, whether an Nrf2 activator can protect tubules from proteinuria-induced tubular damage via anti-inflammatory and antioxidative stress mechanisms is unknown. Using an Institute of Cancer Research-derived glomerulonephritis (ICGN) mouse model of nephrosis, we examined the effects of dihydro-CDDO-trifluoroethyl amide (dh404), a rodent-tolerable bardoxolone methyl analog, in protecting the tubulointerstitium; dh404 markedly suppressed tubular epithelial cell damage in the renal interstitium of ICGN mice. The tubular epithelial cells of ICGN mice showed a decrease in the size and number of mitochondria, as well as the breakdown of the crista structure, whereas the number and ultrastructure of mitochondria were maintained by the dh404 treatment. To further determine the effect of dh404 on mitochondrial function, we used human proximal tubular cells in vitro. Stimulation with albumin and free fatty acid increased mitochondrial reactive oxygen species (ROS). However, dh404 administration diminished mitochondrial ROS. Our data show that dh404 significantly reduced proteinuria-induced tubular cell mitochondrial damage, suggesting that improved redox balance and mitochondrial function and suppression of inflammation underlie the cytoprotective mechanism of Nrf2 activators, including bardoxolone methyl, in diabetic kidney disease.-Nagasu, H., Sogawa, Y., Kidokoro, K., Itano, S., Yamamoto, T., Satoh, M., Sasaki, T., Suzuki, T., Yamamoto, M., Wigley, W. C., Proksch, J. W., Meyer, C. J., Kashihara, N. Bardoxolone methyl analog attenuates proteinuria-induced tubular damage by modulating mitochondrial function.


Posttraumatic Stress Disorder in Primary Care: A Study of General Practices in England.
Posttraumatic Stress Disorder (PTSD) may be a common issue in primary care in the UK, but there have been no studies of all-cause PTSD in general samples of attenders in this country. The current paper thus explores the extent and distribution of probable PTSD among patients attending general practices in England. Cross-sectional survey data from adult patients (n = 1058) attending 11 general practices in southwest England were analysed. Patients were recruited from waiting rooms and completed anonymous questionnaires, including measures of depression, anxiety and risky alcohol use. Current probable PTSD was measured using the 4-item Primary Care PTSD Scale (PC-PTSD). Results indicated 15.1% of patients that exhibited probable PTSD (PC-PTSD ≥ 3), with higher levels observed in practices from deprived areas. There were 53.8% of patients with probable PTSD that expressed the desire for help with these issues. The analyses suggested that rates were lowest among older adults, and highest among patients who were not in cohabitating relationships or were unemployed. Measures of anxiety and depression were associated with 10-fold and 16-fold increases in risk of probable PTSD, respectively, although there were no discernible associations with risky drinking. Such preliminary findings highlight the need for vigilance for PTSD in routine general practice in the UK, and signal a strong need for additional research and attention in this context.


Neuronal components of the superior and inferior tentacles in the terrestrial slug, Limax marginatus.
To identify the types of neurons and to infer the patterns of connectivity in slug tentacles, we stained the neurons in the superior and inferior tentacles in the terrestrial slug, Limax marginatus, by backfilling of the tentacular nerves with Lucifer yellow. Four types of stained neurons, '(1) sensory neurons', '(2) gamma cells', '(3) ganglion cells', '(4) lateral cells', were identified both in the superior and inferior tentacles. Three subtypes of the sensory neurons, '(1a) round sensory neurons', '(1b) spindle-shaped sensory neurons', and '(1c) small sensory neurons', were found in the digits. The gamma cells and the ganglion cells were interneurons. Three subtypes of gamma cells, '(2a) round monopolar gamma cells', '(2b) round bipolar gamma cells', and '(2c) large gamma cells', were present in the digits. The ganglion cells were composed of '(3a) monopolar ganglion cells', '(3b) bipolar ganglion cells', and '(3c) elongated ganglion cells'. The monopolar and bipolar types were located both in the tentacular ganglia and digits, whereas the elongated type was present only in the tentacular ganglia. The lateral cells, whose function is unknown, were found in the dermo-muscular sheaths of the tentacles. Our study provides the first description of the neuronal map of inferior tentacles in gastropods. The results showed no differences in the morphological features of stained neurons between the superior and inferior tentacles in L. marginatus.


Is inspiratory muscle training effective for individuals with cervical spinal cord injury? A qualitative systematic review.
To perform a systematic review to determine the effect of inspiratory muscle training (IMT) in adults with cervical spinal cord injury (CSCI). A systematic search of the literature on IMT and CSCI according to the Cochrane Collaboration protocol was performed. We searched electronic databases up to August 2003 including MEDLINE and CINAHL, searched reference lists from pertinent articles and books, made personal contact with authors, and hand searched targeted journals to identify potential studies for inclusion. Inclusion criteria for the review included randomized controlled trials published in English comparing IMT with another comparison group among adults with CSCI. Two reviewers abstracted relevant data from included studies. Methodological quality of the studies was assessed using criteria developed by Jadad et al. We also assessed whether the comparison groups were similar at baseline and whether an intention-to-treat analysis was performed. Forty articles were retrieved and three met the inclusion criteria. All studies used inspiratory resistance muscle trainers for at least 15 min, twice daily, five to seven days per week for six to eight weeks. Meta-analysis could not be performed due to differences in study design and outcomes. Only one study reported a positive effect of IMTcompared to control for measures of dyspnoea and pulmonary function. Literature on the effect of IMT among adults with CSCI is scarce and an overall effect could not be confirmed.


A new graphic for quality adjusted life years (Q-TWiST) survival analysis: the Q-TWiST plot.
One of the challenges of interpreting a Quality-adjusted time without symptoms of disease and toxicity (Q-TWiST) analysis is examining the sensitivity of conclusions that may be drawn to varying values of the utility coefficients for days with toxicity and days after disease progression. We present a graphic that parsimoniously displays the impact on median Q-TWiST survival across treatment groups of varying values of the utility coefficients. The goal of the graphic is to present a concise Q-TWiST analysis. We use Zhao and Tsiatis (Biometrika 1997; 84(2): 339-348) to adjust for the bias in Kaplan-Meier (K-M) estimates. The graphic contains bounds that approximate points for which statistical significance would be achieved by comparing the median Q-TWiST survival between treatment alternatives for each value of the utility coefficients. The plot may be generalized to compare Q-TWiST means, medians or percentiles across treatment groups. We demonstrate the application of the Q-TWiST plot through a re-analysis of a randomized phase III North Central Cancer Treatment Group (NCCTG) clinical trial of recombinant Interferon-2alpha in patients with malignant melanoma. We explore alternative options to customize the graphic representation for other data sets drawn from several NCCTG clinical trials.


Tetralogy of Fallot with congenital aortic valvar stenosis: the tetralogy-truncus interrelationship.
Two rare patients are reported with tetralogy of Fallot and congenital aortic valvar stenosis. The anatomic and developmental interrelationship between tetralogy of Fallot and truncus arteriosus is summarized. A study of 100 randomly selected postmortem cases of tetralogy revealed aortic valve pathology in 8%, myxomatous aortic valve leaflets without stenosis in 4%, bicuspid aortic valves without stenosis in 3%, and congenital aortic valvar stenosis in 1%. The frequency of systemic semilunar valve pathology in truncus was much higher (66%): moderate to marked myxomatous change in 44%, mild myxomatous change in 22%, truncal valvar stenosis in 11%, and truncal valvar regurgitation in 15%. Being aware of the tetralogy-truncus interrelationship and knowing that myxomatous aortic valves are prone to premature calcific aortic stenosis and/or regurgitation, physicians should follow the aortic valves of surgically repaired patients with tetralogy of Fallot and truncus arteriosus long term with great care. Timely aortic valvuloplasty or replacement may well prove life-saving in such patients.


Interhemispheric connections between olfactory bulbs improve odor detection.
Interhemispheric connections enable interaction and integration of sensory information in bilaterian nervous systems and are thought to optimize sensory computations. However, the cellular and spatial organization of interhemispheric networks and the computational properties they mediate in vertebrates are still poorly understood. Thus, it remains unclear to what extent the connectivity between left and right brain hemispheres participates in sensory processing. Here, we show that the zebrafish olfactory bulbs (OBs) receive direct interhemispheric projections from their contralateral counterparts in addition to top-down inputs from the contralateral zebrafish homolog of olfactory cortex. The direct interhemispheric projections between the OBs reach peripheral layers of the contralateral OB and retain a precise topographic organization, which directly connects similarly tuned olfactory glomeruli across hemispheres. In contrast, interhemispheric top-down inputs consist of diffuse projections that broadly innervate the inhibitory granule cell layer. Jointly, these interhemispheric connections elicit a balance of topographically organized excitation and nontopographic inhibition on the contralateral OB and modulate odor responses. We show that the interhemispheric connections in the olfactory system enable the modulation of odor response and contribute to a small but significant improvement in the detection of a reproductive pheromone when presented together with complex olfactory cues by potentiating the response of the pheromone selective neurons. Taken together, our data show a previously unknown function for an interhemispheric connection between chemosensory maps of the olfactory system.


From nanographene and graphene nanoribbons to graphene sheets: chemical synthesis.
Graphene, an individual two-dimensional, atomically thick sheet of graphite composed of a hexagonal network of sp(2) carbon atoms, has been intensively investigated since its first isolation in 2004, which was based on repeated peeling of highly oriented pyrolyzed graphite (HOPG). The extraordinary electronic, thermal, and mechanical properties of graphene make it a promising candidate for practical applications in electronics, sensing, catalysis, energy storage, conversion, etc. Both the theoretical and experimental studies proved that the properties of graphene are mainly dependent on their geometric structures. Precise control over graphene synthesis is therefore crucial for probing their fundamental physical properties and introduction in promising applications. In this Minireview, we highlight the recent progress that has led to the successful chemical synthesis of graphene with a range of different sizes and chemical compositions based on both top-down and bottom-up strategies.


A translational inhibitor from muscles of diabetic rats: identification as histone H1.
A heat- and acid-stable protein fraction that inhibited peptide chain initiation in rabbit reticulocyte lysates was extracted from frozen, powdered rat skeletal muscles by stepwise trichloroacetic acid precipitation. Streptozotocin-induced diabetes increased the inhibitory activity; this was prevented by insulin therapy. Size-exclusion high-performance liquid chromatography resolved four inhibitory fractions; only one was consistently increased (approximately 2-fold) in muscle extracts from diabetic rats. Polysome profiles of lysates incubated with this fraction indicated peptide chain initiation inhibition. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified inhibitory fraction migrated with apparent Mr 30 and 32 kDa, which on Western blot immunostained with antisera against histone H1/H1(0). Perchloric acid extraction of muscle homogenates yielded approximately twofold more H1 from diabetic than from control rats; yield from diabetics decreased to control values 5 h after subcutaneous insulin injection. Inclusion of detergent during homogenization increased H1 yield more from muscles of control than from diabetic rats and abolished the difference between them. Because H1 affects several biochemical reactions, its facilitated extraction from insulin-deprived tissues can bias interpretation of studies of insulin action.


Reverse-transcriptional gene expression of anammox and ammonia-oxidizing archaea and bacteria in soybean and rice paddy soils of Northeast China.
The relative gene expression of hydrazine oxidoreductase encoding gene (hzo) for anaerobic ammonium oxidizing bacteria (anammox) and ammonia monooxygenase encoding gene (amoA) for both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in Sanjiang Plain soybean and rice paddy soils of Northeast China was investigated by using real-time reverse-transcriptional quantitative PCR. Metabolically active populations of anammox, AOA, and AOB in rice paddy soils were evident by the presence and successful quantification of hzo mRNA and amoA mRNA genes. The expression ratio of amoA gene for both AOA and AOB varied between soybean soils and different rice paddy soils while the expression of hzo gene for anammox was detectable only in rice paddy soils by showing a diverse relative expression ratio in each soil sample. Gene expression of both archaeal and bacterial amoA genes in rice paddy soils differed among the three sampling depths, but that of hzo was not. Both archaeal and bacterial amoA genes showed an increase trend of expression level with continuation of rice paddy cultivation, but the low expression ratio of hzo gene indicated a relatively small contribution of anammox in overall removal of inorganic nitrogen through N2 even under anoxic and high nitrogen input in agriculture. Bacterial amoA gene from two soybean fields and three rice paddy fields were also analyzed for community composition by denaturing gradient gel electrophoresis fingerprint. Community shift was observed between soybean and paddy fields and within each of them. The consistent occurrence of three bands 5, 6, and 7 in all samples showed their high adaptability for both arid cultivation and continuous rice paddy cultivation. Our data suggest that AOA and AOB are playing a more important role in nitrogen transformation in agricultural soils in oxic or anoxic environment and anammox bacteria may also contribute but in a less extent to N transformation in these agricultural soils under anoxic condition.


Modeling epidemics dynamics on heterogenous networks.
The dynamics of the SIS process on heterogenous networks, where different local communities are connected by airlines, is studied. We suggest a new modeling technique for travelers movement, in which the movement does not affect the demographic parameters characterizing the metapopulation. A solution to the deterministic reaction-diffusion equations that emerges from this model on a general network is presented. A typical example of a heterogenous network, the star structure, is studied in detail both analytically and using agent-based simulations. The interplay between demographic stochasticity, spatial heterogeneity and the infection dynamics is shown to produce some counterintuitive effects. In particular it was found that, while movement always increases the chance of an outbreak, it may decrease the steady-state fraction of sick individuals. The importance of the modeling technique in estimating the outcomes of a vaccination campaign is demonstrated.


Role of magnesium chelatase activity in the early steps of the tetrapyrrole biosynthetic pathway.
Magnesium-protoporphyrin IX chelatase (Mg-chelatase) is located at the branchpoint of tetrapyrrole biosynthesis, at which point protoporphyrin IX is distributed for the synthesis of chlorophyll and heme. We investigated the regulatory contribution of Mg-chelatase to the flow of metabolites. In plants, the enzyme complex consists of three subunits, designated CHL D, CHL I, and CHL H. Transgenic tobacco (Nicotiana tabacum) plants expressing antisense RNA for the Mg-chelatase subunit CHL H were analyzed to elucidate further the role of Mg-chelatase in the distribution of protoporphyrin IX into the branched tetrapyrrolic pathway. The transgenic plants displayed a reduced growth rate and chlorophyll deficiency. Both phenotypical properties were correlated with lower Mg-chelatase activity. Unexpectedly, less protoporphyrin IX and heme accumulated, and a decrease in 5-aminolevulinate (ALA)-synthesizing capacity and ALA dehydratase activity paralleled the progressive reduction in Mg-chelatase activity in the transformants compared with control plants. The reduced activities of the early enzymatic steps corresponded with lower levels of transcripts encoding glutamyl-tRNA reductase and ALA-dehydratase. The decreased expression and activities of early enzymes in the pathway could be explained by a feedback-controlled mechanism in response to lower Mg-chelatase activity. We discuss intercompartmental signaling that synchronizes the activities of the first steps in tetrapyrrolic metabolism with the late steps for the synthesis of end products.


NetB and necrotic enteritis: the hole movable story.
Clostridium perfringens is the primary causative agent of avian necrotic enteritis. Our understanding of the pathogenesis of this economically important disease has been enhanced by the discovery of C. perfringens NetB toxin, which belongs to the α-haemolysin family of β-pore-forming toxins. In a chicken disease model, the analysis of an isogenic set of strains comprising the wild type, a netB mutant, and its complemented derivative, fulfilled molecular Koch's postulates and revealed that NetB was essential for disease. These results were consistent with epidemiological surveys, which generally found that there was a higher prevalence of netB carriage in C. perfringens isolates from diseased poultry compared to healthy birds. The netB gene has been shown to be located on large conjugative plasmids that are closely related to other toxin plasmids from C. perfringens, which has potential implications for the epidemiology of necrotic enteritis infections. The crystal structures of both monomeric NetB and the heptameric NetB pore have been determined, the latter revealed a central pore diameter of approximately 26 Å. Finally, it has been shown that vaccine preparations that include NetB can protect chickens against disease and a series of single amino acid substitution derivatives of NetB that have potential value for vaccine formulations have been isolated and analysed. It is likely that NetB will be an important antigen to include in an effective, commercially viable, necrotic enteritis vaccine.


The effect of oxandrolone on body proportions and body composition in growth hormone-treated girls with Turner syndrome.
Untreated girls with Turner syndrome (TS) have short stature, relatively broad shoulders, a broad pelvis, short legs, a high fat mass and low muscle mass. Our objective was to assess the effect of the weak androgen oxandrolone (Ox) on body proportions and composition in growth hormone (GH)-treated girls with TS. 133 patients were included in a randomized, placebo-controlled, double-blind study. Patients were treated with GH (1.33 mg/m(2) per day) from baseline, combined with placebo (Pl) or Ox in a low (0.03 mg/kg per day) or previously conventional (0.06 mg/kg per day) dose from the age of eight, and oestrogens from the age of twelve. Sitting height, biacromial and biiliacal distances compared with height (i.e. shape values), BMI, waist circumference, sum of 4 skinfolds (sum4skin) and upper arm muscle area (UAMA) SD scores (SDS) were assessed half-yearly. Compared with GH + Pl, adult shape values on GH + Ox tended to be higher for sitting height (Ox 0.03, P = 0.2; Ox 0.06, P = 0.02) and biacromial distance (Ox 0.03, P = 0.2; Ox 0.06, P = 0.07) and lower for biiliacal distance (Ox 0.03, P = 0.004; Ox 0.06, P = 0.08). Sum4skin SDS tended to decrease more (Ox 0.03, P = 0.2; Ox 0.06, P = 0.005) while UAMA SDS increased more (Ox 0.03, P < 0.001; Ox 0.06, P < 0.001) than on GH + Pl. The increase in BMI and waist circumference SDS was comparable between the dosage groups. In GH-treated girls with TS, Ox 0.06 increases sitting height and tends to increase biacromial distance and decrease biiliacal distance, while Ox 0.03 significantly decreases biiliacal distance compared with height. Furthermore, Ox 0.06 reduces subcutaneous fat mass, and both Ox dosages increase muscle mass.


Occurrence and fate of perfluorinated compounds in sewage sludge from Spain and Germany.
Perfluorinated compounds (PFCs) are persistent and bioaccumulative organic compounds used as additives in many industrial products. After use, these compounds enter wastewater treatment plants (WWTP) and long-chain PFCs are primarily accumulated in sludge. The aim of this study was to determine the occurrence and behavior of five PFCs in sludge from 15 WWTP from Spain and Germany that receive both urban and industrial wastes. The PFCs studied were perfluorooctanesulfonate (PFOS), perfluorohexanesulfonate (PFHxS), perfluorobutanesulfonate (PFBS), perfluorooctanoic acid (PFOA), and perfluorononanoic acid (PFNA). One gram of freeze-dried, sieved, and homogenized sludge was extracted using an ultrasonic bath with methanol and glacial acetic acid. After that, the extract was recovered and evaporated to dryness with a TurboVap and then 1 mL of acetonitrile was added and the extract was cleaned up with black carbon. Liquid chromatography coupled to mass spectrometry operated in selected reaction monitoring was used to determine target compounds. Quality parameters are provided for the set of compounds studied. PFCs were detected in all samples. In Spanish sludge, ∑PFC ranged from 0.28 to 5.20 ng/g dry weight (dw) with prevalence of PFOS, while in German sludge, ∑PFC ranged from 20.7 to 38.6 ng/g dw and PFBS was the dominant compound. As a next step, the evolution of PFC concentrations within the sludge treatment steps (primary sludge, anaerobic digested sludge, and centrifuged sludge) was evaluated and differences among levels and patterns were observed and were attributed to the influent water quality and treatment used. Finally, we estimated the amount of PFCs discharged via sludge in order to determine the potential impact to the environment according to different sludge usage practices in the two regions investigated. This manuscript provided an intra-European overview of PFC distribution in sludge. Levels and compound distribution depend on the WWTP sampled. This study demonstrates that PFCs are persistent to sludge treatment and the loads in sludge may pose a future environmental risk, if not controlled.


Mechanism-Based Inhibitors of the Human Sirtuin 5 Deacylase: Structure-Activity Relationship, Biostructural, and Kinetic Insight.
The sirtuin enzymes are important regulatory deacylases in a variety of biochemical contexts and may therefore be potential therapeutic targets through either activation or inhibition by small molecules. Here, we describe the discovery of the most potent inhibitor of sirtuin 5 (SIRT5) reported to date. We provide rationalization of the mode of binding by solving co-crystal structures of selected inhibitors in complex with both human and zebrafish SIRT5, which provide insight for future optimization of inhibitors with more "drug-like" properties. Importantly, enzyme kinetic evaluation revealed a slow, tight-binding mechanism of inhibition, which is unprecedented for SIRT5. This is important information when applying inhibitors to probe mechanisms in biology.


Effect of genetic variations in syntaxin-binding protein-5 and syntaxin-2 on von Willebrand factor concentration and cardiovascular risk.
Elevated von Willebrand factor (VWF) plasma levels are associated with an increased risk of cardiovascular disease. A meta-analysis of genomewide association studies on VWF identified novel candidate genes, that is, syntaxin-binding protein 5 (STXBP5) and syntaxin 2 (STX2), which are possibly involved in the secretion of VWF. We investigated whether VWF antigen levels (VWF:Ag), VWF collagen-binding activity (VWF:CB) and the risk of arterial thrombosis are affected by common genetic variations in these genes. In 463 young white subjects (men ≤45 years of age and women ≤55 years of age), who were included 1 to 3 months after a first event of arterial thrombosis, and 406 control subjects, we measured VWF:Ag and VWF:CB. Nine haplotype tagging single-nucleotide polymorphisms of STXBP5 and STX2 were selected and subsequently analyzed using linear regression with additive genetic models adjusted for age, sex, and ABO blood group. The minor alleles of rs9399599 and rs1039084 in STXBP5 were associated with lower VWF plasma levels and activity, whereas the minor allele of rs7978987 in STX2 was associated with higher VWF plasma levels and activity. The minor alleles of the single-nucleotide polymorphisms in STX2 were associated with a reduced risk of arterial thrombosis (rs1236: odds ratio, 0.73 [95% confidence interval, 0.59, 0.89]; rs7978987: odds ratio, 0.81 [95% confidence interval, 0.65, 1.00]; rs11061158: odds ratio, 0.69 [95% confidence interval, 0.55, 0.88]). Genetic variability in STXBP5 and STX2 affects both VWF concentration and activity in young individuals with premature arterial thrombosis. Furthermore, in our study, genetic variability in STX2 is associated with the risk of arterial thrombosis. However, at this point, the underlying mechanism remains unclear.


A member of the ALOG gene family has a novel role in regulating nodulation in Lotus japonicus.
Legumes can control the number of symbiotic nodules that form on their roots, thus balancing nitrogen assimilation and energy consumption. Two major pathways participate in nodulation: the Nod factor (NF) signaling pathway which involves recognition of rhizobial bacteria by root cells and promotion of nodulation, and the autoregulation of nodulation (AON) pathway which involves long-distance negative feedback between roots and shoots. Although a handful of genes have a clear role in the maintenance of nodule number, additional unknown factors may also be involved in this process. Here, we identify a novel function for a Lotus japonicus ALOG (Arabidopsis LSH1 and Oryza G1) family member, LjALOG1, involved in positively regulating nodulation. LjALOG1 expression increased substantially after inoculation with rhizobia, with high levels of expression in whole nodule primordia and in the base of developing nodules. The ljalog1 mutants, which have an insertion of the LORE1 retroelement in LjALOG1, had significantly fewer nodules compared with wild type, along with increased expression of LjCLE-RS1 (L. japonicus CLE Root Signal 1), which encodes a nodulation suppressor in the AON pathway. In summary, our findings identified a novel factor that participates in controlling nodulation, possibly by suppressing the AON pathway.


Characterizing systemic exposure of inhaled drugs: application to the long-acting β2-agonist PF-00610355.
PF-00610355 is an orally inhaled long-acting β2-adrenoreceptor agonist that is being developed for the once-daily treatment of chronic obstructive pulmonary disease (COPD). The pharmacological effect is exerted in the lungs. However, systemic exposure of PF-00610355 is expected to be responsible for certain drug-related adverse effects. This analysis characterizes PF-00610355 using an integrated analysis of systemic exposure, across trials and patient populations. A total of 6,107 samples of PF-00610355 plasma concentration, collected in 264 subjects from eight studies in healthy volunteers, asthma, and COPD patients, were analyzed using non-linear mixed-effects models. Model-based mean (95 % CI) exposure profiles for a range of PF-00610355 doses in COPD patients were simulated. PF-00610355 exposure profiles were described by a three-compartment disposition model with first-order absorption through a transit compartment. Patient status, inhalation device, and demographic factors were found to influence systemic drug exposure. Relative fine particle dose had a minor effect, whereas no effect of baseline lung function on the systemic exposure was found. An implicit method to address pharmacokinetic variability between occasions of drug intake yielded similar results as the established explicit method, yet in a much more efficient way. The estimated systemic pre-dose and maximum PF-00610355 plasma concentration was 23 and 38 % in COPD patients compared to healthy volunteers, respectively. The analysis illustrated the value of an integrated pharmacokinetic analysis to address specific challenges in the clinical development of long-/ultra-long-acting β2-agonists and inhaled compounds in general, both in relation to selecting a safe starting dose in patients, but also in understanding exposure and systemic safety information across different patient populations and different inhalation devices/formulations.


Cardiovascular collapse in infants: association with paroxysmal atrial tachycardia.
Four infants, aged 16 to 28 days (mean 23 days), were seen in the emergency room with acute cardiovascular collapse and with normal heart rate and rhythm. During evaluation for cardiovascular collapse, no infant had sepsis; cardiac assessment revealed normal intracardiac anatomy but global cardiac chamber enlargement and poor left ventricular systolic function, which resolved with supportive treatment. However, three of the four infants demonstrated ventricular preexcitation on their surface electrocardiogram and, subsequently, two infants had transient episodes of tachycardia. During a transesophageal pacing study to evaluate inducibility and electrophysiologic characteristics of tachycardia, sustained tachycardia was initiated in all four infants. Reentrant tachycardia used an accessory atrioventricular connection as evidenced by the presence of preexcitation during sinus rhythm (three infants), the ability to initiate and terminate tachycardia by programmed electrical stimulation (four infants), minimum ventriculoatrial interval recorded in the esophagus (V-Aeso) exceeded 70 ms (four infants), transient bundle branch block during tachycardia prolonged the cycle length and the V-Aeso by 30 to 50 ms (three infants). Findings in these infants suggested prior episodes of prolonged tachycardia as the probable etiology of the cardiovascular collapse.


Limitation of physical performance in a muscle fatiguing handgrip exercise is mediated by thalamo-insular activity.
In this study, we investigated central/supraspinal processes mediating cessation of a muscle fatiguing exercise. Fifteen male subjects performed 39 intermittent, isometric handgrip contractions (13 s on, 5-6 s off) with the dominant right hand while brain activation was assessed by means of functional magnetic resonance imaging (fMRI). An adaptive, partly stochastic protocol was designed such that in approximately 50% of the contraction trials the required force could not be held until the end of the trial (task failure trial). Trials performed in compliance with the force requirements (succeeded trial) were compared with task failure trials concerning neural activity during a small time window before task failure occurred. The data revealed significantly increased activation contralaterally in both the mid/anterior insular cortex and the thalamus during the investigated time window in the case of subsequent task failure. In accordance with other studies investigating sensations that alert the organism to urgent homeostatic imbalance such as air hunger, hunger for food, and pain, we assume that an increased thalamo-insular activation in the context of a fatigue-induced handgrip exercise could reflect increased homeostatic disturbance in the exercising muscle and may be of essential importance by mediating task failure to maintain the integrity of the organism.


Optimization of folate-conjugated liposomal vectors for folate receptor-mediated gene therapy.
A folate-targeted transfection complex that is internalized by certain cancer cells and displays several properties reminiscent of enveloped viruses has been developed. These liposomal vectors are comprised of a polycation-condensed DNA plasmid associated with a mixture of neutral and anionic lipids supplemented with folate-poly(ethylene glycol)-dioleylphosphatidylethanolamine for tumor cell-specific targeting. N-Citraconyl-dioleylphosphatidylethanolamine is also included for pH-dependent release of endosome-entrapped DNA into the cytoplasm, and a novel plasmid containing a 366-bp segment from SV40 DNA has been employed to facilitate transport of the plasmid into the nucleus. Because formation of the DNA core is an important step in the assembly of liposomal vectors, considerable effort was devoted to comparing the transfection efficiencies of various DNA condensing agents. It was found that complexation of plasmid DNA with high molecular weight polymers such as acylated-polylysine and cationic dendrimers leads to higher folate-mediated transfection efficiency than DNA complexed with unmodified polylysine. In contrast, compaction of plasmid DNA with small cationic molecules such as spermine, spermidine, or gramicidin S yields only weakly active folate-targeted liposomal vectors. Compared to analogous liposomal vector preparations lacking an optimally compacted DNA core, a cell-specific targeting ligand, a caged fusogenic lipid, and a nucleotide sequence that facilitates nuclear uptake, these modified liposomal vectors display greatly improved transfection efficiencies and target cell specificity.


Distribution and role in regeneration of N-CAM in the basal laminae of muscle and Schwann cells.
The neural cell adhesion molecule (N-CAM) is a membrane glycoprotein involved in neuron-neuron and neuron-muscle adhesion. It can be synthesized in various forms by both nerve and muscle and it becomes concentrated at the motor endplate. Biochemical analysis of a frog muscle extract enriched in basal lamina revealed the presence of a polydisperse, polysialylated form of N-CAM with an average Mr of approximately 160,000 as determined by SDS-PAGE, which was converted to a form of 125,000 Mr by treatment with neuraminidase. To define further the role of N-CAM in neuromuscular junction organization, we studied the distribution of N-CAM in an in vivo preparation of frog basal lamina sheaths obtained by inducing the degeneration of both nerve and muscle fibers. Immunoreactive material could be readily detected by anti-N-CAM antibodies in such basal lamina sheaths. Ultrastructural analysis using immunogold techniques revealed N-CAM in close association with the basal lamina sheaths, present in dense accumulation at places that presumably correspond to synaptic regions. N-CAM epitopes were also associated with collagen fibrils in the extracellular matrix. The ability of anti-N-CAM antibodies to perturb nerve regeneration and reinnervation of the remaining basal lamina sheaths was then examined. In control animals, myelinating Schwann cells wrapped around the regenerated axon and reinnervation occurred only at the old synaptic areas; new contacts between nerve and basal lamina had a terminal Schwann cell capping the nerve terminal. In the presence of anti-N-CAM antibodies, three major abnormalities were observed in the regeneration and reinnervation processes: (a) regenerated axons in nerve trunks that had grown back into the old Schwann cell basal lamina were rarely associated with myelinating Schwann cell processes, (b) ectopic synapses were often present, and (c) many of the axon terminals lacked a terminal Schwann cell capping the nerve-basal lamina contact area. These results suggest that N-CAM may play an important role not only in the determination of synaptic areas but also in Schwann cell-axon interactions during nerve regeneration.


Animal welfare concerns during the use of the water bath for stunning broilers, hens, and ducks.
European legislation demands that slaughter animals, including poultry, be rendered immediately unconscious and insensible until death occurs through blood loss at slaughter. This study addressed requirements for stunner settings (i.e., voltage, wave oscillation frequency) and response parameters (i.e., applied current, behavior) affecting effective water bath stunning. An inventory of current electrical stunning practice was performed in 10 slaughterhouses in the Netherlands. Thereafter, measurements were performed using a single-bird water bath to examine the effects of stunner settings based on the average technical settings observed in the slaughterhouses. Responses were recorded at 50, 400, and 1,000 Hz on broilers and hens and at 50 and 400 Hz on ducks under controlled laboratory conditions. Effects of voltage settings (broilers: 100 to 400 V; hens: 150 to 300 V; ducks: 150 to 400 V) on current levels (broilers: 45 to 444 mA; hens: 40 to 219 mA; ducks: 64 to 362 mA) and consciousness (response to pain stimulus) were recorded immediately after stunning. Brain and heart activity was monitored using electroencephalogram and electrocardiogram technology. Results show that effective stunning using the conventional water bath almost exclusively produces blood splashing in broilers. Effective stunning current levels did not differ significantly between broilers, hens, and ducks effectively stunned hens tended to require lower currents. Effective stuns at higher frequencies resulted in higher currents. Similar input voltage (V) levels (within and between bird type) resulted in significant variation (P < 0.001) in current levels (mA) required for an effective stun, indicating variability in electrical impedance between individual birds. Body weight and bird type did not affect the probability of an effective stun. Multi-bird water bath usage does not ensure effective stunning and technical adjustments can result in detrimental effects on meat quality. Future legislation should consider wave form, relationships between frequency and current allowing for individual impedance variation and effects on meat quality while safeguarding animal welfare.


Fusion expression of green fluorescent protein and HCV capsid antigene in Escherichia coli cells.
A chimeric gene of the green fluorescent protein (GFP) and hepatitis C virus (HCV) core antigene were constructed and expressed in E. coli cells. The expressed fusion protein was examined by Dot-ELISA and Western blot and the three antigenic determinants were detected. The GFP-Core fusion protein showed not only the striking green fluorescence under natural light but also the HCV antigenic activity. A new method of immunological diagnosis is greatly anticipated in the light of this fusion protein which can be seen as the HCV antigen tagged with the green fluorescent protein.


Naturally Occurring TPE-CA Maintains Gut Microbiota and Bile Acids Homeostasis via FXR Signaling Modulation of the Liver-Gut Axis.
Antibiotics-induced changes in intestinal flora (dysbiosis) may have various effects on the host. Dysbiosis is associated with numerous metabolites including bile acids, which are produced in the liver from cholesterol and metabolized in the gut by intestinal microbiota. Total phenolic extracts of Citrus aurantium L. (TPE-CA) are rich in dietary flavanones and their glycosyl derivatives, including flavones, flavonols, polymethoxyflavones and coumarins, which exert positive health effects on the microbiota. The aim of this study is to elucidate the interplays between the intestinal microbiota and bile acids metabolism attributed to antibiotics. Mice were exposed to broad-spectrum antibiotics, such as ampicillin, streptomycin and clindamycin, for 14 days. This exposure resulted in reduced bacterial diversity and richness, and destroyed intestinal permeability. The homeostasis of bile acids was also affected. Subsequent TPE-CA administration, counteracted most of the dysbiosis, and reshaped intestinal permeability, these effects occurred via upregulation of zonula occludens 1 and occludin associated proteins and downregulation of serum endotoxin compared to the antibiotics group. TPE-CA maintained the homeostasis of bile acids via modulation of the liver-gut axis related farnesoid X receptor (FXR)/fibroblast growth factor 15 (FGF15) pathway and FXR-targeted protein. Our findings indicated that TPE-CA exerted a protective effect on the restoration of intestinal microbiota composition, reshaped barrier integrity and maintained bile acid homeostasis via the liver-gut axis with antibiotics-induced dysbiosis.


Modulation of the reactivity of the thiol of human serum albumin and its sulfenic derivative by fatty acids.
The single cysteine residue of human serum albumin (HSA-SH) is the most abundant plasma thiol. HSA transports fatty acids (FA), a cargo that increases under conditions of diabetes, exercise or adrenergic stimulation. The stearic acid-HSA (5/1) complex reacted sixfold faster than FA-free HSA at pH 7.4 with the disulfide 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and twofold faster with hydrogen peroxide and peroxynitrite. The apparent pK(a) of HSA-SH decreased from 7.9±0.1 to 7.4±0.1. Exposure to H(2)O(2) (2mM, 5min, 37°C) yielded 0.29±0.04mol of sulfenic acid (HSA-SOH) per mole of FA-bound HSA. The reactivity of HSA-SOH with low molecular weight thiols increased ∼threefold in the presence of FA. The enhanced reactivity of the albumin thiol at neutral pH upon FA binding can be rationalized by considering that the corresponding conformational changes that increase thiol exposure both increase the availability of the thiolate due to a lower apparent pK(a) and also loosen steric constraints for reactions. Since situations that increase circulating FA are associated with oxidative stress, this increased reactivity of HSA-SH could assist in oxidant removal.


Enabling Nurse-Patient Communication With a Mobile App: Controlled Pretest-Posttest Study With Nurses and Non-English-Speaking Patients.
There is growing concern regarding the implications of miscommunication in health care settings, the results of which can have serious detrimental impacts on patient safety and health outcomes. Effective communication between nurses and patients is integral in the delivery of timely, competent, and safe care. In a hospital environment where care is delivered 24 hours a day, interpreters are not always available. In 2014, we developed a communication app to support patients' interactions with allied health clinicians when interpreters are not present. In 2017, we expanded this app to meet the needs of the nursing workforce. The app contains a fixed set of phrases translated into common languages, and communication is supported by text, images, audio content, and video content. This study aims to evaluate the efficacy of the communication app to support nursing staff during the provision of standard care to patients from non-English-speaking backgrounds when an interpreter is not available. This study used a one-group pretest-posttest sequential explanatory mixed methods research design, with quantitative data analyzed using inferential statistics and qualitative data analyzed via thematic content analysis. A total of 134 observation sessions (82 pretest and 52 posttest) of everyday nurse-patient interactions and 396 app use sessions were recorded. In addition, a total of 134 surveys (82 pretest and 52 posttest) with nursing staff, 7 interviews with patients, and 3 focus groups with a total of 9 nursing staff participants were held between January and November 2017. In the absence of the app, baseline interactions with patients from English-speaking backgrounds were rated as more successful (t80=5.69; P<.001) than interactions with patients from non-English-speaking backgrounds. When staff used the app during the live trial, interactions with patients from non-English-speaking backgrounds were rated as more successful than interactions without the app (F2,119=8.17; P<.001; η2=0.37). In addition, the level of staff frustration was rated lower when the app was used to communicate (t80=2.71; P=.008; r=0.29). Most participants indicated that the app assisted them in communicating. Through the use of the app, a number of patients from non-English-speaking backgrounds experienced better provision of standard care, similar to their English-speaking peers. Thus, the app can be seen as contributing to the delivery of equitable health care.


Catheter ablation of atrial fibrillation in patients with hypertrophic cardiomyopathy: atrial fibrillation type determines the success rate.
Atrial fibrillation (AF) in patients with hypertrophic cardiomyopathy (HCM) is generally associated with deterioration of the clinical status, functional capacity, and quality of life. It is also an independent risk factor for stroke and death. Studies evaluating the effectiveness of AF ablation in this cohort are relatively scant, have included relatively few patients, and their results are somewhat conflicting. Thus, the aim of this study was to assess the safety and efficacy of catheter ablation of AF in patients with HCM. Thirty patients (10 females; mean age 48.7 ± 11 years) with drug-refractory paroxysmal (n = 14), persistent (n = 7), or long-persistent (> 1 year; n = 9) AF were prospectively recruited into the study. Eleven patients were in New York Heart Association (NYHA) class I, 13 patients were in NYHA class II, and 6 patients were in NYHA class III. Mean atrial volume was 180 ± 47 mL, interventricular septum thickness was 20.5 ± 6.3 mm, and left atrial area was 29.8 ± 6.2 cm2. Ablation protocol was adjusted to the clinical and electrophysiological status of the patients. Pulmonary vein isolation and bidirectional cavo-tricuspid isthmus block were performed in all patients. In addition, left atrial linear lesions were created and complex fragmented atrial potentials were ablated in patients with persistent and long-persistent AF, as well as during repeated procedures. At 12 months, stable sinus rhythm (SR) was present in 16 (53%) patients, significantly more frequently in patients with paroxysmal AF (71% in SR) compared to those with persistent (57.1% in SR) or long-persistent (22% in SR) AF. A significant reduction of AF burden was observed in 85.7% of patients with paroxysmal AF, 71.4% of patients with persistent AF, and 55.5% of patients with long-persistent AF. Single procedure success rate was 33% (10 patients), and repeat ablation procedures were performed in 13 patients. No periprocedural complications occurred. Thromboembolic events were noted in 2 patients with arrhythmia recurrence during the follow-up, including stroke in 1 patient and peripheral embolism in the other patient. In both these patients, heart failure worsening was observed during these events, and anticoagulation was inadequate in one of them. Five of 16 patients in whom stable SR was observed during the follow-up were off antiarrhythmic drug therapy at final evaluation. In the other 6 patients, antiarrhythmic drug therapy was continued due to ventricular arrhythmias. Successfully treated patients more often had paroxysmal AF (successful ablation: paroxysmal AF in 10 of 16 patients; unsuccessful ablation: paroxysmal AF in 4 of 14 patients; p = 0.009) and were younger (45 ± 11.5 years vs. 52.6 ± 9.2 years; p = 0.046). In addition, a trend toward a reduced need for cardioversion at the end of the procedure was also observed in these patients (3 patients in the successful ablation group vs. 8 patients in the unsuccessful ablation group; p = 0.056). In multivariate regression analysis, paroxysmal AF was the only independent predictor of a successful outcome. Catheter ablation of AF in patients with HCM is an effective and safe therapeutic option, particularly in patients with paroxysmal AF. Effectiveness of ablation is significantly smaller in patients with persistent AF and even more so in those with long-persistent AF. Repeated procedures were often necessary. Continued antiarrhythmic drug therapy is often required due to a significant degree of atrial remodelling.


Influenza-Like Illness in Travelers to the Developing World.
Travelers to developing regions are at risk for development of influenza-like illness (ILI). Little is known of traveler and trip characteristics associated with the development of ILI. TravMil is a prospective observational study, enrolling subjects presenting to six military travel clinics or predeployment-screening sites. We analyzed pre- and post-travel surveys from travelers visiting regions outside of the continental United States, Western or Northern Europe, Canada, Australia, or New Zealand between January 2010 and March 2016. Influenza-like illness was defined as a self-reported fever associated with either sore throat or cough. Trip and traveler characteristics were analyzed to determine risk factors for the development of ILI. Two thousand nine hundred and thirty-two trips were recorded (55% male, median age 45 years, 69% white, 51% on vacation, median travel duration 17 days). The 2,337 trips included the number of self-reported influenza vaccinations in the preceding 5 years (median 5). Eleven percent of the trips were complicated by an ILI lasting a median of 5 days; 70% and 17% of these reported upper and lower respiratory tract infection, respectively, and 12% reported both. On multivariate analysis, increased risk of ILI was associated with female gender (odds ratio [OR]: 1.60 [confidence interval (CI): 1.25-2.05], P < 0.01), age (years) (OR: 1.01 [CI: 1.01-1.02], P < 0.01); and duration of travel (days) (OR: 1.01 [CI: 1.00-1.01], P < 0.01). Influenza-like illness is common in travelers, regardless of traveler characteristics, purpose of travel, destination, or season of year. Female gender, older age, and longer duration of travel were associated with an increased risk of ILI. Additional tools and strategies are needed to prevent ILI in international travelers.


Transcriptional regulation of the human UDP-galactose:ceramide galactosyltransferase (hCGT) gene expression: functional role of GC-box and CRE.
UDP-galactose:ceramide galactosyltransferase (CGT, EC 18.104.22.168) is a key enzyme in the biosynthetic pathway of galactocerebroside (GalC), the most abundant glycolipid in myelin. Using a GalC expressing cell line, human oligodendroglioma (HOG), one which does not express GalC, human neuroblastoma (LAN-5), we previously demonstrated that the human CGT (hCGT) gene promoter functions in a cell-specific manner. Because the proximal (-292/-256) and distal (-747/-688) positive domains were shown to be critically involved in regulating the expression of several myelin-specific genes, we further investigated the functional roles of these two motifs in hCGT expression. Mutation analysis confirmed that a GC-box (-267/-259) and a CRE (-697/-690) were critical for hCGT expression. Electrophoretic mobility shift assay (EMSA) demonstrated that these motifs specifically bound to nuclear extracts from both cell lines. Using antibodies to Sp1, Sp3, pCREB-1, and ATF-1, these proteins were shown to be components of the EMSA complexes. However, the only difference between the HOG and LAN-5 cells was found in the EMSA profile of the CRE complexes. This difference may account for the differential transcription of the hCGT gene in the two cell types. Furthermore, the expression levels of ATF-1 detected were much higher in HOG cells than in LAN-5 cells. Thus, our data suggest that the GC-box and CRE function cooperatively, and that the CRE regulates the cell-specific expression of the hCGT gene.


A comparison of long-term outcome in first-episode schizophrenia following treatment with risperidone or a typical antipsychotic.
Most reports assessing the efficacy and tolerability of risperidone have involved patients previously treated with typical antipsychotics. Such patients are more likely to have a greater resistance or intolerance to treatment, thus restricting our interpretation of the impact a new treatment might have on the course of schizophrenia and possibly biasing the results. The present study examines the relative effectiveness of risperidone and typical antipsychotics in patients being treated for their first episode of schizophrenia. From a cohort of 126 patients, 2 groups of 19 first-episode DSM-III-R/DSM-IV schizophrenia patients matched for age, gender, length of illness, and length of treatment and treated with either a typical antipsychotic or risperidone for a minimum of 1 year were compared on a number of outcome dimensions during their course of treatment and at follow-up. Treatment allocation was not random, and patients were judged to be compliant with medication. Patients treated with typical antipsychotics were followed up for a statistically nonsignificantly longer time (mean = 2.7 vs. 1.9 years). Six patients (31.6%) from the typical antipsychotic group were admitted to the hospital within the first year following the index admission compared with 1 patient (5.3%) in the risperidone group (admitted at month 14). Patients in the risperidone group showed a statistically significantly lower length of first hospitalization (p < .01), utilization of inpatient beds during the course of treatment (p < .001), and use of anticholinergic medication (p < .05). There were no statistically significant differences in symptom levels, either during the course of treatment or at follow-up; in the use of antidepressant, antianxiety, or mood-stabilizing drugs; or in changes in living circumstances or employment. These findings confirm at least equal long-term efficacy of typical antipsychotics and risperidone, but a possible advantage for risperidone in decreased service utilization and decreased use of anticholinergic drugs.


Retinal vein occlusion in child with rare mutations in genes for thrombophilia.
Retinal vein occlusions (RVOs) are rare in the younger population. Hematological pro-thrombotic factors are thought to be important in a minority, amplifying an atherosclerotic anatomical predisposition. Anotherwise healthy 13-year-old girl presented two episodes of sudden decreased vision in few months.Ophthalmological exams pointed out post-thrombotic intra-retinal hemorrhage. All investigations were normal, thrombophilia screen showed factor XII deficiency and genetic mutations of methylenetetrahydrofolate reductase (MTHFR), angiotesin convertin enzyme (ACE) and angiotensinogen (AGT). Two intravitreal injection of bevacizumab was administered with improving visual acuity; subsequently the patient did not report further episodes. In addition to traditional factors with procoagulant activity, factor XII deficiency plays an important role in thrombosis's mechanism. Its deficiency causes a marked prolongation of the activated partial thromboplastin time in the laboratory examination. Moreover also MTHFR, ACE and AGT could have been involved in this case, so it is important to evaluate these parameters in the differential diagnosis of RVOs.


Survival analysis of the association between antenatal care attendance and neonatal mortality in 57 low- and middle-income countries.
Neonatal mortality is unacceptably high in most low- and middle-income countries (LMICs). In these countries, where access to emergency obstetric services is limited, antenatal care (ANC) utilization offers improved maternal health and birth outcomes. However, evidence for this is scanty and mixed. We explored the association between attendance for ANC and survival of neonates in 57 LMICs. Employing standardized protocols to ensure comparison across countries, we used nationally representative cross-sectional data from 57 LMICs (N = 464 728) to investigate the association between ANC visits and neonatal mortality. Cox proportional hazards multivariable regression models and meta-regression analysis were used to analyse pooled data from the countries. Kaplan-Meier survival curves were used to describe the patterns of neonatal survival in each region. After adjusting for potential confounding factors, we found 55% lower risk of neonatal mortality [hazard ratio (HR) 0.45, 95% confidence interval (CI) 0.42-0.48] among women who met both WHO recommendations for ANC (first visit within the first trimester and at least four visits during pregnancy) in pooled analysis. Furthermore, meta-analysis of country-level risk shows 32% lower risk of neonatal mortality (HR 0.68, 95% CI 0.61-0.75) among those who met at least one WHO recommendation. In addition, ANC attendance was associated with lower neonatal mortality in all the regions except in the Middle East and North Africa. ANC attendance is protective against neonatal mortality in the LMICs studied, although differences exist across countries and regions. Increasing ANC visits, along with other known effective interventions, can improve neonatal survival in these countries.


Separation of Tc-99 in soil and plant samples collected around the chernobyl reactor using a Tc-selective chromatographic resin and determination of the nuclide by ICP-MS
Technetium (Tc) is known to have high mobility in a soil-water system and also high bioavailability for plants, because the most stable form of Tc in natural surface environment is thought to be TcO4- which is highly soluble. The chemical form of Tc, however, changes with environmental conditions. Thus, it is necessary, for realistic assessment, to obtain transfer parameters, such as transfer factors, under natural conditions. However, it is difficult to obtain these parameters using global fallout 99Tc in actual fields due to its low concentration. In this study, 99Tc concentrations in surface soil and plant leaf samples collected from forest sites within the 30-km zone around the Chernobyl reactor were measured for the first time. In the case of soil samples, a simple and rapid analytical method for determination of 99Tc is used which consists of volatilizing and trapping Tc in a combustion apparatus, purifying the Tc with an extraction chromatographic resin, and measuring it by inductively coupled plasma mass spectrometry (ICP-MS). For plant samples, a wet digestion method in combination with the resin is applied and the 99Tc is measured by ICP-MS. Concentrations of 99Tc in organic soil samples and leaves of strawberry (Fragaria vesca) range from 1.1-14.8 Bq kg(-1) dry weight and 0.2-6.0 Bq kg(-1) dry weight, respectively. Our results indicate that soil-to-plant transfer factors for Tc are similar to those for Cs.


Overcoming PGP-related multidrug resistance. The cyclosporine derivative SDZ PSC 833 can abolish the resistance to methoxy-morpholynil-doxorubicin.
The results obtained so far in studies designed to neutralize P glycoprotein (PGP)-related multidrug resistance (MDR) by using MDR reversal agents, have not yet fulfilled the promise of the experiments which were performed in vitro. In order to improve PGP-related MDR neutralization, we tested in vitro the activity of the cyclosporine derivative SDZ PSC 833 (PSC) together with doxorubicin (DOX) and with two new DOX derivatives named 4' iodo 4' deoxy-doxorubicin (IODODOX) and methoxy-morpholynil-doxorubicin (MMDOX, FCE 23762) using four different human cell lines and their multi-drug resistant variants. Anthracycline toxicity was evaluated by using the MTT method after a 7-day culture with continuous exposure to the antitumor drugs with or without the addition of PSC. PSC significantly downmodulated the toxicity of all three anthracyclines in all the four cell systems. However, despite the great increase caused by PSC in the toxicity of DOX and a more modest effect on the toxicity of the two DOX derivatives, this MDR reversal agent could only completely block the PGP mediated MMDOX resistance whereas DOX refractoriness was only decreased. The combination of MMDOX or IODODOX with PSC 1.6 microM is more efficient than the combination of DOX plus PSC for the full reversion of PGP-mediated drug resistance. Careful clinical studies are required to evaluate if these associations can also effectively and safely neutralize MDR in vivo.


Selection for intrinsic endurance modifies endocrine stress responsiveness.
Physical exercise dampens an individual's stress response and decreases symptoms of anxiety and depression disorders. While the extrinsic relationship of exercise and psychological state is established, their intrinsic relationship is unresolved. We investigated the potential intrinsic relationship of exercise with stress responsiveness using NIH rats bidirectionally selected for intrinsic endurance capacity. Selection resulted in two populations, one with high intrinsic endurance (high capacity runners; HCR) and one with low intrinsic endurance (low capacity runners; LCR). Animals from these populations were subjected to the elevated plus maze (EPM) and novel environment to assess levels of anxiety-like behavior, and to restraint stress to determine stress responsiveness. Pre-test plasma corticosterone levels and the response of plasma corticosterone to exposure to the EPM and restraint were analyzed using ELISA. A dexamethasone suppression test was performed to assess negative feedback tone of corticosterone release. Pre-test plasma corticosterone levels were similar between LCR and HCR, and these populations had similar behavioral and corticosterone responses to the EPM. Following restraint, HCR animals exhibited more anxiotypic behavior than LCR animals on the EPM, and exhibited an increase in plasma corticosterone following EPM and restraint that was not observed in LCR animals. HCR animals also exhibited more anxiotypic behavior in the novel environment compared to LCR animals. Plasma corticosterone levels were equally reduced in both populations following dexamethasone administration. Overall, our data suggest a positive genetic relationship between exercise endurance and stress responsiveness, which is at odds with the established extrinsic relationship of these traits.


Essential role of brc-2 in chromosome integrity of germ cells in C. elegans.
brc-2, an ortholog of BRCA2 in Caenorhabditis elegans, is essential in the maintenance of genetic integrity. In C. elegans, cellular location correlates with meiotic progression, and transgene-induced cosuppression is observed in the germ line but not in somatic cells. We used these unique features to dissect the role of brc-2 in the germ line from that in somatic cells. In situ hybridization of wild type animals revealed that brc-2 gene expression was higher in oocytes than in other germline cells, and was barely detectable in mitotic cells. In contrast, germ cells containing multicopies of the brc-2 transgene showed no significant in situ hybridization signal at any oogenesis stage, confirming that brc-2 expression was functionally cosuppressed in the transgenic germ line. RAD-51 foci formation in response to DNA damage was abrogated in brc-2-cosuppressed germ cells, whereas wild-type germ cells showed strong RAD-51 foci formation. These germ cells exhibited massive chromosome fragmentation and decompaction instead of six bivalent chromosomes in diakinesis. Accordingly, lethality was observed after the early stage of germline development. These results suggest that brc-2 plays essential roles in chromosome integrity in early prophase, and therefore is crucial in meiotic progression and embryonic survival.


Study of exciton transfer in dense quantum dot nanocomposites.
Nanocomposites of colloidal quantum dots (QDs) integrated into conjugated polymers (CPs) are key to hybrid optoelectronics, where engineering the excitonic interactions at the nanoscale is crucial. For such excitonic operation, it was believed that exciton diffusion is essential to realize nonradiative energy transfer from CPs to QDs. In this study, contrary to the previous literature, efficient exciton transfer is demonstrated in the nanocomposites of dense QDs, where exciton transfer can be as efficient as 80% without requiring the assistance of exciton diffusion. This is enabled by uniform dispersion of QDs at high density (up to ∼70 wt%) in the nanocomposite while avoiding phase segregation. Theoretical modeling supports the experimental observation of weakly temperature dependent nonradiative energy transfer dynamics. This new finding provides the ability to design hybrid light-emitting diodes that show an order of magnitude enhanced external quantum efficiencies.


The Evolutionary Dynamics of a Genetic Barrier to Gene Flow: From the Establishment to the Emergence of a Peak of Divergence.
Divergent selection works when an allele establishes in the subpopulations in which it is adaptive, but not in the ones in which it is deleterious. While such a locally adaptive allele is maintained, the target locus of selection works as a genetic barrier to gene flow or a barrier locus. The genetic divergence (or FST) around the barrier locus can be maintained, while in other regions of the genome, genetic variation can be mixed by gene flow or migration. In this work, we consider theoretically the evolutionary process of a barrier locus, from its birth to stable preservation. Under a simple two-population model, we use a diffusion approach to obtain analytical expressions for the probability of initial establishment of a locally adaptive allele, the reduction of genetic variation due to the spread of the adaptive allele, and the process to the development of a sharp peak of divergence (genomic island of divergence). Our results will be useful to understanding how genomes evolve through local adaptation and divergent selection.


[Marker chromosomes of murine cell lines].
Using the C-method of chromosome staining four marker chromosomes were revealed in the transplanted murine line SC-1, one comparatively rare marker chromosome was shown in RAG line, small marker chromosomes occurred almost in all cells of RVP3 line. Marker chromosomes found in the studied lines by the C-method of chromosome staining make it possible to distinguish these lines from each other.


Dislocation filtering in GaN nanostructures.
Dislocation filtering in GaN by selective area growth through a nanoporous template is examined both by transmission electron microscopy and numerical modeling. These nanorods grow epitaxially from the (0001)-oriented GaN underlayer through the approximately 100 nm thick template and naturally terminate with hexagonal pyramid-shaped caps. It is demonstrated that for a certain window of geometric parameters a threading dislocation growing within a GaN nanorod is likely to be excluded by the strong image forces of the nearby free surfaces. Approximately 3000 nanorods were examined in cross-section, including growth through 50 and 80 nm diameter pores. The very few threading dislocations not filtered by the template turn toward a free surface within the nanorod, exiting less than 50 nm past the base of the template. The potential active region for light-emitting diode devices based on these nanorods would have been entirely free of threading dislocations for all samples examined. A greater than 2 orders of magnitude reduction in threading dislocation density can be surmised from a data set of this size. A finite element-based implementation of the eigenstrain model was employed to corroborate the experimentally observed data and examine a larger range of potential nanorod geometries, providing a simple map of the different regimes of dislocation filtering for this class of GaN nanorods. These results indicate that nanostructured semiconductor materials are effective at eliminating deleterious extended defects, as necessary to enhance the optoelectronic performance and device lifetimes compared to conventional planar heterostructures.


Three-dimensional solution structure of the HIV-1 protease complexed with DMP323, a novel cyclic urea-type inhibitor, determined by nuclear magnetic resonance spectroscopy.
The three-dimensional solution structure of the HIV-1 protease homodimer, MW 22.2 kDa, complexed to a potent, cyclic urea-based inhibitor, DMP323, is reported. This is the first solution structure of an HIV protease/inhibitor complex that has been elucidated. Multidimensional heteronuclear NMR spectra were used to assemble more than 4,200 distance and angle constraints. Using the constraints, together with a hybrid distance geometry/simulated annealing protocol, an ensemble of 28 NMR structures was calculated having no distance or angle violations greater than 0.3 A or 5 degrees, respectively. Neglecting residues in disordered loops, the RMS deviation (RMSD) for backbone atoms in the family of structures was 0.60 A relative to the average structure. The individual NMR structures had excellent covalent geometry and stereochemistry, as did the restrained minimized average structure. The latter structure is similar to the 1.8-A X-ray structure of the protease/DMP323 complex (Chang CH et al., 1995, Protein Science, submitted); the pairwise backbone RMSD calculated for the two structures is 1.22 A. As expected, the mismatch between the structures is greatest in the loops that are disordered and/or flexible. The flexibility of residues 37-42 and 50-51 may be important in facilitating substrate binding and product release, because these residues make up the respective hinges and tips of the protease flaps. Flexibility of residues 4-8 may play a role in protease regulation by facilitating autolysis.


Integration and safety of fertility preservation in a breast cancer program.
Young women diagnosed with breast cancer typically face systemic treatments that may delay childbearing or permanently impair their fertility. These concerns add to the stress experienced by young cancer survivors. Timely counseling and providing fertility preservation through cryopreservation of eggs or embryos have become an important quality of life issue. We analyzed the impact of fertility preservation procedures on the initiation of treatment for breast cancer and discuss critical aspects of the process.


Blood-brain barrier dysfunction underlying Alzheimer's disease is induced by an SSAO/VAP-1-dependent cerebrovascular activation with enhanced Aβ deposition.
Dysfunctions of the vascular system directly contribute to the onset and progression of Alzheimer's disease (AD). The blood-brain barrier (BBB) shows signs of malfunction at early stages of the disease. When Abeta peptide (Aβ) is deposited on brain vessels, it induces vascular degeneration by producing reactive oxygen species and promoting inflammation. These molecular processes are also related to an excessive SSAO/VAP-1 (semicarbazide-sensitive amine oxidase) enzymatic activity, observed in plasma and in cerebrovascular tissue of AD patients. We studied the contribution of vascular SSAO/VAP-1 to the BBB dysfunction in AD using in vitro BBB models. Our results show that SSAO/VAP-1 expression is associated to endothelial activation by altering the release of pro-inflammatory and pro-angiogenic angioneurins, most highly IL-6, IL-8 and VEGF. It is also related to a BBB structure alteration, with a decrease in tight-junction proteins such as zona occludens or claudin-5. Moreover, the BBB function reveals increased permeability and leukocyte adhesion in cells expressing SSAO/VAP-1, as well as an enhancement of the vascular Aβ deposition induced by mechanisms both dependent and independent of the enzymatic activity of SSAO/VAP-1. These results reveal an interesting role of vascular SSAO/VAP-1 in BBB dysfunction related to AD progression, opening a new window in the search of alternative therapeutic targets for fighting AD.


Arthroscopic trans-portal deep medial collateral ligament pie-crusting release.
Arthroscopic treatments of meniscal injuries of the knee are among the most common orthopaedic procedures performed. Adequate visualization of the posterior horn of the medial meniscus might be challenging, especially in patients with tight medial compartments. In these cases instrument manipulation in an attempt to reach the posterior horn of the meniscus can cause an iatrogenic chondral injury because of the narrow medial joint space. A transcutaneous medial collateral ligament (MCL) pie-crusting release facilitates expansion of the medial joint space in a case of a tight medial compartment. Nevertheless, it might cause injury to the superficial MCL, infection, and pain and injury to the saphenous nerve because of multiple needle punctures of the skin. We describe an inside-out, arthroscopic deep MCL pie-crusting release, which allows access to the medial meniscus through the anterior approach to provide good visualization of the footprint and sufficient working space.


Comparison between young and old patients with bronchogenic carcinoma.
The aim of this study was to compare the clinical characteristics, histological type, stage at diagnosis, treatment and survival of young (< or = 45 years) and older patients with bronchogenic carcinoma. The study was designed as a retrospective review of all lung cancer patients referred to the Kuwait Cancer Control Center over a 10-year period from 1985 to 1994. The study comprised 590 patients with primary bronchogenic carcinoma, of whom 72 (12%) were < or = 45 years of age at the time of diagnosis. Median (range) duration of symptoms, percentage of smokers and male to female ratios for the younger and older patients were 8 (2-48) weeks vs. 8 (1-52) weeks (p = 0.9), 74% vs. 83% (p = 0.06) and 5.5 vs. 5.3, respectively. The majority of patients had advanced stage disease at presentation; 91% of the younger patients had stage III or IV compared with 88% of the older patients (p = 0.1). The histological types for the younger and older patients were 32% vs. 20% for adenocarcinoma (p = 0.01) and 33% vs. 45% for squamous cell carcinoma (p = 0.1). There were more patients in the younger group who had surgery (21% vs. 7.5%) than in the older group (p = 0.001). Follow-up data were available for 177 patients out of 190 Kuwaiti national patients (93%). The median (range) survival rates for young and old patients were 8 (3-62) months and 7 (1-174) months, respectively (p = 0.09). Only 6 patients survived for more than 5 years, one (7.6%) from the younger group and 5 (3%) from the older group. Our study shows a relatively high percentage of young patients in our population of lung cancer patients. Apart from the higher incidence of adenocarcinoma in the young group and the fact that these patients had more surgery than the older group, there were no other significant differences between the two groups.


Allelopathic Potential of Invasive Plantago virginica on Four Lawn Species.
Plantago virginica L. has invaded many lawn ecosystems in the Eastern part of China. The invasion has incurred an economic cost to remove them. In order to prevent the invasion, it is critical to understand the invasive mechanisms of this species. However, few studies have been conducted on the allelopathic mechanisms of its invasion. In this study, we examined allelopathic effects of P. virginica on germination of seeds and growth of seedlings of four widely used lawn species. We found extensive allelopathic potential of P. virginica on other lawn species, which varied with species and developmental stage. While most effects of the extracts of P. virginica were inhibitory, some variables in some species were promoted by the addition of the extracts. The extracts of P. virginica significantly inhibited seed germination of Agrostis matsumurae. While the overall differences in seed germination rate of Poa annua were significant among treatments, difference between control and any of the treatments was not significant. The height of seedlings of A. matsumurae and Cynodon dactylon was significantly lower under the treatments of adding extracts of P. virginica. In contrast, growth of seedlings of Festuca elata and P. annua did not show significant differences among treatments. The root length of A. matsumurae, C. dactylon and P. annua was suppressed by the extracts of P. virginica whereas root length of F. elata was not affected. Aboveground biomass of A. matsumurae and F. elata was significantly higher than control, except for F. elata at the concentration of 50mg/mL, whereas aboveground biomass of C. dactylon and P. annua was reduced at higher concentrations of the extracts. Except for A. matsumurae, root biomass of the other three lawn species declined under the treatments with the extracts of P. virginica. Our results revealed that P. virginica had allelopathic potential on four lawn species and supported the theory of "novel weapons hypothesis". Invasion by P. virginica in lawn can be moderated by selecting those species that are not affected or promotionally affected by it.


Allergy to laboratory animals in children of parents occupationally exposed to mice, rats and hamsters.
Sensitization to laboratory animals (LA) has a high prevalence among laboratory workers. It is unknown whether transportation of LA allergens can be a risk factor for sensitization of subjects outside the laboratory environment. The aim of the study was to investigate the prevalence of sensitization to LA among children whose parents were and were not occupationally exposed to LA. The first group consisted of 50 children (age 12.3+/-4.3 yrs) whose parents were occupationally exposed to mice, rats and hamsters. The second group consisted of 40 children (age (mean+/-SD) 10.8+/-3.0 yrs) whose parents were not occupationally exposed to LA. Children having LA at home were eliminated from the study. All children responded to a questionnaire, underwent spirometry and were also tested with skin prick tests with the use of common allergens and prick tests with hair extracts from mouse, hamster and rat. Total immunoglobulin (Ig)E levels and the presence of specific IgE against LA were also estimated. Children of parents occupationally exposed to LA presented significantly more positive skin prick tests against allergens from the hair of laboratory animals compared to children of nonexposed parents. Five children from the first group were also found to have specific IgE against LA, with three of these five children complaining of rhinitis and cough while visiting their parents' workplace. It is concluded that the observed increased sensitization to laboratory animals among children of occupationally exposed parents could be the result of poor hygienic conditions at their parents' workplace. Hence, parents' job seems to be an additional risk factor of sensitization and should be taken into consideration when recording an allergic history.


Role of sialic acid in erythrocyte survival.
The role of membrane sialic acid in erythrocyte survival is unclear, although there is evidence for a reduction in sialic acid and surface charge in older erythrocytes. We reduced the surface charge of human, rat, and rabbit erythrocytes by removing sialic acid with neuraminidase. Reduction in sialic acid correlated with decreases in electrophoretic mobility and loss of PAS staining of membrane glycoproteins on polyacrylamide gels. No changes in ATP levels or deformability were found. 51Cr erythrocyte survivals in rats and rabbits showed rapid clearance of desialylated erythrocytes with sequestration by the liver. These results suggest that reduction in erythrocyte sialic acid is a mechanism of erythrocyte destruction and may be important in erythrocyte senescence.


Effect of weight reduction on liver histology and biochemistry in patients with chronic hepatitis C.
Steatosis occurs in more than 50% of patients with chronic hepatitis C and is associated with increased hepatic fibrosis. In many of these patients the pathogenesis of steatosis appears to be the same as for patients with non-alcoholic fatty liver disease-that is, related to visceral adiposity and obesity. The effect of a three month weight reduction programme on liver biochemistry and metabolic parameters was examined in 19 subjects with steatosis and chronic hepatitis C. Paired liver biopsies were performed in 10 subjects, prior to and 3-6 months following the intervention, to determine the effect of weight loss on liver histology. There was a mean weight loss of 5.9 (3.2) kg and a mean reduction in waist circumference of 9.0 (5.0) cm. In 16 of the 19 patients, serum alanine aminotransferase levels fell progressively with weight loss. Mean fasting insulin fell from 16 (7) to 11 (4) mmol/l (p<0.002). Nine of 10 patients with paired liver biopsies had a reduction in steatosis irrespective of viral genotype. In these subjects the median modified Knodell fibrosis score decreased from 3 to 1 (p=0.04) and activated stellate cells significantly decreased (p<0.004). Weight loss in patients with chronic hepatitis C may be associated with a reduction in steatosis and abnormal liver enzymes and an improvement in fibrosis, despite the persistence of the virus. Weight reduction may provide an important adjunct treatment strategy for patients with chronic hepatitis C.


Non-linear cardiac dynamics and morning dip: an unsound circadian rhythm.
The frequency of sudden cardiac death increases in the morning. The relationship between decreased complexity of heart rate dynamics and sudden cardiac death has been documented. An understanding of circadian variation in the complexity of cardiac dynamics may be important to predict and prevent sudden cardiac death. Dynamic 24-h electrocardiographic recordings were obtained from 30 healthy ambulant subjects aged 41-50 years, and the digitized data were partitioned into sections of 30 min duration. For each section, four indexes obtained from separate algorithms of non-linear dynamics of the RR interval--modified correlation dimension, Lyapunov exponent, approximate entropy, and fractal dimension--were calculated. Normalized low-(0.04-0.15 hertz) and high-frequency (> 0.15 hertz) components were also calculated. All four indexes of non-linear dynamics showed a remarkably similar circadian rhythm: a prominent morning dip preceded by a steep decline during the late night, a recovery during the evening and a peak around midnight. In the morning, the low-frequency component rose rapidly with concomitant reduction in the high-frequency component. The complexity of cardiac dynamics decreases significantly in the morning, and this may contribute to the ominously increased rate of cardiac death in the morning hours.


Hypophosphorylated SR splicing factors transiently localize around active nucleolar organizing regions in telophase daughter nuclei.
Upon completion of mitosis, daughter nuclei assemble all of the organelles necessary for the implementation of nuclear functions. We found that upon entry into daughter nuclei, snRNPs and SR proteins do not immediately colocalize in nuclear speckles. SR proteins accumulated in patches around active nucleolar organizing regions (NORs) that we refer to as NOR-associated patches (NAPs), whereas snRNPs were enriched at other nuclear regions. NAPs formed transiently, persisting for 15-20 min before dissipating as nuclear speckles began to form in G1. In the absence of RNA polymerase II transcription, NAPs increased in size and persisted for at least 2 h, with delayed localization of SR proteins to nuclear speckles. In addition, SR proteins in NAPs are hypophosphorylated, and the SR protein kinase Clk/STY colocalizes with SR proteins in NAPs, suggesting that phosphorylation releases SR proteins from NAPs and their initial target is transcription sites. This work demonstrates a previously unrecognized role of NAPs in splicing factor trafficking and nuclear speckle biogenesis.


Telomere length assessment in human archival tissues: combined telomere fluorescence in situ hybridization and immunostaining.
A method was developed to assess human telomere lengths at the individual cell level in tissue sections from standard formalin-fixed paraffin-embedded tissues. We coupled this method with immunofluorescence to allow the simultaneous identification of specific cell types. Validation of this in situ quantification method showed excellent agreement with the commonly used telomere repeat fragment-Southern blot method. The assay requires very few cells ( approximately 10 to 15). Thus, small tissue samples, including clinical biopsies, can be easily accommodated. In addition, the cells under study need not be actively cycling and there is no requirement for tissue disaggregation or cell culture. This method provides a more accurate assessment of telomere lengths than Southern blotting because confounding contributions from undesired cell types within tissue samples are avoided. Using this technique, we were able to perform the first comparison of relative telomere lengths in matched tumor versus normal epithelial cells within archival human prostate tissues.