Synthesis and luminescence studies of mono- and C3-symmetric, tris(ligand) complexes of Sm(III), Y(III) and Eu(III) with sulfur-bridged binaphtholate ligands.
A series of trivalent mono- and tris(ligand) lanthanide complexes of a sulfur-bridged binaphthol ligand [1,1'-S(2-HOC(10)H(4)Bu(t)(2)-3,6)(2)] H(2)L(SN), have been prepared and characterised both structurally and photophysically. The H(2)L(SN) ligand provides an increased steric bulk and offers an additional donor atom (sulfur) as compared with 1,1'-binaphthol (BINOL), a ligand commonly used to complex Lewis acidic lanthanide catalysts. Reaction of the diol H(2)L(SN) with [Sm[N(SiMe(3))(2)](3)] affords silylamido- and amino- derivatives [Sm(L(SN))[N(SiMe(3))(2)][HN(SiMe(3))(2)]] and the crystallographically characterised [Sm(L(SN))[N(SiMe(3))(2)](thf)(2)] with different degrees of structural rigidity, depending on the presence of coordinating solvents. The binaphthyl groups of the L(SN) ligand act as sensitisers of the metal centred emission, which is observed for the Eu(III) and Sm(III) complexes studied. We have therefore sought to use emission spectroscopy as a non-invasive technique to monitor a monomer-dimer equilibrium in these complexes. A dramatic difference between the emission properties of the unreactive dimeric Sm(III) aryloxide complex, the solvated monomeric analogues and the amido adduct demonstrated the potential use of such a technique. For a few representative lanthanides (Ln = Sm, Eu and Y) the reaction of the dilithium salt Li(2)L(SN) with either [Ln[N(SiMe(3))(2]3)] or [LnCl(3)(thf)(3)] affords only the homoleptic complex [Li(S)(3)][LnL(SN)(3)](S = thf or diethyl ether); we report the structural characterisation of the Sm complex. However, the reactions of this dipotassium salt K(2)L(SN) with [Sm[N(SiMe(3))(2)](3)] or [SmCl(3)(thf)(3)] give only [SmL(SN)N(SiMe(3))(2)], or intractable mixtures respectively, in which no (tris)binaphtholate is observed. The only isolable lanthanide-L(SN) halide adduct so far is [YbL(SN)I(thf)].


Field desorption mass spectrometry of large multiply branched saturated hydrocarbons.
Large multiply branched saturated hydrocarbons containing 67-103 carbon atoms (molecular masses 941.8-1446.8 Da) were analyzed by field desorption mass spectrometry (FD-MS) with a double-focusing mass spectrometer. FD-MS was found to have detection limits in the 100 fmol range. The FD mass spectra exhibited molecular ions of astonishingly low abundance. However, the fragment ions formed were closely related to the proposed molecular structure, allowing us to set up rules for straightforward structure elucidation of unknowns. In detail, (i) dehydrogenation, (ii) alkyl losses from molecular ions and (iii) subsequent alkene losses were observed. The influence of the electric field strength on dehydrogenation and C-C cleavages was examined by variation of the emitter potential. Additionally, ion dissociations in the ion source and in the first and second field-free regions, respectively, were compared to study the relative importance of field-induced and thermally induced processes.


Electrochemical Enzyme Biosensor Bearing Biochar Nanoparticle as Signal Enhancer for Bisphenol A Detection in Water.
An electrochemical tyrosinase enzyme (Tyr) biosensor using a highly conductive sugarcane derived biochar nanoparticle (BCNP) as a transducer and signal enhancer (BCNPs/Tyr/Nafion/GCE) was developed for the sensitive detection of bisphenol A (BPA). The BCNPs/Tyr/Nafion/GCE biosensor exhibited improved amperometric current responses such as higher sensing signal, decreased impedance and lowered reduction potential compared with the Tyr/Nafion/GCE due to high conductivity property of the biochar nanoparticle. Under the optimized conditions, it could detect BPA in good sensitivity with linear range from 0.02 to 10 μM, and a lowest detection limit of 3.18 nM. Moreover, it showed a low Km value, high reproducibility and good selectivity over other reagents, and the BCNPs/Tyr complex solution also showed good stability with 86.9% of sensing signal maintained after one month storage. The biosensor was also successfully utilized for real water detection with high accuracy as validated by high performance liquid chromatography. Therefore, the biochar nanoparticle based enzyme biosensor proved to be a potential and reliable method for high performance detection of pollutants in the environment.


Unusual features of primary dermatofibrosarcoma protuberans and its myxoid recurrence.
A case of Dermatofibrosarcoma Protuberans (DFSP) with unusual clinical and histological features is presented. The lesion recurred one year after surgical removal and showed myxoid degeneration of the entire lesion. Only a few cases with myxoid degeneration of the entire lesion are described in the literature. We found CD 34 stain, in combination with a panel of antibodies including fXIIIa, S100 and actin, useful in the diagnosis of this type of tumor in order to differentiate it from other neural, fibrous or muscular neoplasms.


Evaluation of amenorrhea.
Pregnancy is the most common cause of amenorrhea and must be ruled out before proceeding with diagnostic evaluation. A careful history and physical examination may reveal evidence of androgen excess, estrogen deficiency or other endocrinopathies. Serum prolactin and thyroid-stimulating hormone (TSH) levels should be checked in all women who are not pregnant. Galactorrhea by history or on examination and/or an elevated prolactin level should be investigated with an imaging study to rule out a pituitary adenoma. If serum prolactin and TSH levels are normal, a progesterone challenge test should be performed to determine outflow tract patency and estrogen status. In women with hypoestrogenic amenorrhea, indicated by a negative challenge test and a competent outflow tract, serum gonadotropin, follicle-stimulating hormone and luteinizing hormone levels may be measured to determine whether amenorrhea represents ovarian failure or pituitary or hypothalamic dysfunction. Hypothalamic amenorrhea is common in women with a history of weight loss, stress or vigorous exercise. Amenorrheic women with adequate estrogen levels should receive cyclic progesterone. Hormonal therapy and calcium supplementation in hypoestrogenic amenorrhea.


Quaternary epitope specificities of anti-HIV-1 neutralizing antibodies generated in rhesus macaques infected by the simian/human immunodeficiency virus SHIVSF162P4.
Monoclonal antibodies (MAbs) that neutralize human immunodeficiency virus type 1 (HIV-1) have been isolated from HIV-1-infected individuals or animals immunized with recombinant HIV-1 envelope (Env) glycoprotein constructs. The epitopes of these neutralizing antibodies (NAbs) were shown to be located on either the variable or conserved regions of the HIV-1 Env and to be linear or conformational. However, one neutralizing MAb, 2909, which was isolated from an HIV-1-infected subject, recognizes a more complex, quaternary epitope that is present on the virion-associated functional trimeric Env spike of the SF162 HIV-1 isolate. Here, we discuss the isolation of 11 anti-HIV NAbs that were isolated from three rhesus macaques infected with the simian/human immunodeficiency virus SHIV(SF162P4) and that also recognize quaternary epitopes. A detailed epitope mapping analysis of three of these rhesus antibodies revealed that their epitopes overlap that of the human MAb 2909. Despite this overall similarity in binding, however, differences in specific amino acid and glycosylation pattern requirements for MAb 2909 and the rhesus MAbs were identified. These results highlight similarities in the B-cell responses of humans and macaques to structurally complex neutralization epitopes on related viruses, HIV-1 and SHIV.


Pseudomonas aeruginosa clinical isolates: serotypes, resistance phenotypes and plasmid profiles.
78 Pseudomonas aeruginosa strains were isolated from the respiratory tract of 56 patients, 15 of which were affected by cystic fibrosis (CF). The epidemiological typing scheme was based on serotyping, antibiotic resistance pattern and plasmid DNA profile. All strains (except 2 mucoid strains) were typed using a rapid slide O-agglutination technique. Most common serotypes in both group were 0:1, 0:10 and 0:6. Moreover we observed a correlation among 0:12 serotype and CF patients. Plasmid DNA analysis showed that 45.2% (on average) of strains isolated from patients with and w/o CF harboured 1-3 plasmids ranging in size from 1 to 15 Md. Plasmid prevalence was higher in strains isolated from CF patients in specimens collected after antibiotic therapy. A correlation was found between 1 and 1.9 Md plasmids and resistance to aminoglycosides. Our results indicate that the analysis of antibiotic resistance phenotypes combined with plasmid analysis may be useful, in association to serotyping, to characterize the circulation of P. aeruginosa strains and the spread of resistance in these bacteria.


Ontology-aware neural network: a general framework for pattern mining from microbiome data.
With the rapid accumulation of microbiome data around the world, numerous computational bioinformatics methods have been developed for pattern mining from such paramount microbiome data. Current microbiome data mining methods, such as gene and species mining, rely heavily on sequence comparison. Most of these methods, however, have a clear trade-off, particularly, when it comes to big-data analytical efficiency and accuracy. Microbiome entities are usually organized in ontology structures, and pattern mining methods that have considered ontology structures could offer advantages in mining efficiency and accuracy. Here, we have summarized the ontology-aware neural network (ONN) as a novel framework for microbiome data mining. We have discussed the applications of ONN in multiple contexts, including gene mining, species mining and microbial community dynamic pattern mining. We have then highlighted one of the most important characteristics of ONN, namely, novel knowledge discovery, which makes ONN a standout among all microbiome data mining methods. Finally, we have provided several applications to showcase the advantage of ONN over other methods in microbiome data mining. In summary, ONN represents a paradigm shift for pattern mining from microbiome data: from traditional machine learning approach to ontology-aware and model-based approach, which has found its broad application scenarios in microbiome data mining.


"What makes some rats live so long?" The mitochondrial contribution to longevity through balance of mitochondrial dynamics and mtDNA content.
Extremely interesting for aging research are those individuals able to reach older ages still with functions similar to those of younger counterparts. We examined liver samples from ad libitum-fed old (28-month-old, AL-28) and ad libitum-fed very old (32-month-old, AL-32) rats for a number of markers, relevant for mitochondrial functionality and mitochondrial DNA (mtDNA) content. As for the mtDNA content and the protein amounts of the citrate synthase and the antioxidant peroxiredoxin III there were no significant changes in the AL-32 animals. No significant longevity-related change was found for TFAM amount, but a 50% reduction in the amount of the Lon protease, responsible for turnover of TFAM inside mitochondria, characterized the AL-32 rats. No longevity-related change was observed also for the amounts of the mtDNA repair enzymes OGG1 and APE1, whereas the intra-mitochondrial amount of the cytochrome c protein showed a 50% increase in the AL-32 rats, indicating a likely reduced initiation of the intrinsic apoptotic pathway. Totally unexpected was the doubling of two proteins, very relevant for mitochondrial dynamics, namely MFN2 and DRP1, in the AL-32 rats. This prompted us to the calculation of all individual fusion indexes that grouped together in the AL-32 rats, while in the AL-28 animals were very different. We found a strong positive correlation between the fusion indexes and the respective mtDNA contents in two AL-28 and four AL-32 rats. This supports the idea that the limited prevalence of fusion above a still active fission should have ensured a functional mitochondrial network and should have led to a quite narrow range of high mtDNA contents, likely the best-suitable for extended longevity. Our findings strongly suggest that, among the multiple causes leading to the longevity of the AL-32 rats, the maintenance of an adult-like balance of mitochondrial dynamics seems to be very relevant for the regulation of mtDNA content and functionality.


[Mycobacteria in the environment].
Mycobacteria typically are free-living environmental saprophytes and strict pathogens such as M. tuberculosis and M. leprae, which do not proliferate in the inanimate environment form rare exceptions within the genus Mycobacterium. Many free-living mycobacteria such as M. avium and M. kansasii have been described as potential human pathogens and there is evidence that they are usually transmitted by inhalation, inoculation and ingestion from environmental sources. Soil, bogs, surface water, ground water and sea water have been described as natural habitats of environmental mycobacteria. In addition, artificial habitats such as tap water and sewage sludge are colonized by a considerable spectrum of mycobacterial species. Aquatic mycobacteria tend to colonize biofilms at air-water and solid-water interfaces and the latter seem to be an important proliferation site in oligotrophic habitats. Outside the host tissue M. chelonae ssp. abscessus, M. kansasii, M. simiae, and M. xenopi have so far almost exclusively been isolated from artificial oligotrophic aquatic habitats, i.e. tap water and drinking water; their natural reservoir is unknown. Some species have not yet been isolated from the environment although epidemiological data suggest that infections caused by these organisms are acquired from environmental sources.


Seepage Characteristics of Mixed-Wettability Porous Media on the Phase-Field Model.
Clarifying the microscale gas-water flow behaviors in a mixed wettability reservoir is of great importance for underground engineering. A numerical model of mixed wettability based on circular particles was constructed using the MATLAB stochastic distribution program, and the gas-water flow was simulated based on the phase-field method. The Navier-Stokes equations were solved by the finite element method. The work analyzed the effects of the content of heterogeneous wetting particles, wettability, and inversed wettability of the matrix on the flow path and pressure distribution of the mixed wettability model. Besides, the two-phase flow behaviors were evaluated in microscale mixed-wettability porous media. The simulation results revealed that (i) the residual saturation of the gas phase showed a positive correlation with the hydrophobic particle content, and closed gases only existed in isolated pore channels with small content. Isolated closed gases gradually connected as the content increased. (ii) The residual gas content in the corner and tail end increased as the hydrophobicity of particles increased in hydrophilic matrices. Hydrophobic matrices showed a negative correlation, with the greatest pressure drop due to capillary resistance and step changes in the neutral-hydrophobic transition zone. (iii) Water-phase breakthrough time and gas-phase residual saturation showed a negative correlation change. The more space occupied by the gas phase, the faster the water-phase breakthrough. Moreover, the saturation no longer changes after the breakthrough. The work provides a guideline for determining the dominant flow path of phase displacements and the distribution of residual phases.


Single-dose ciprofloxacin versus 3 days of norfloxacin in uncomplicated urinary tract infections in women.
A double-blind, randomized controlled study to evaluate the efficacy and safety of a single dose of 500 mg of ciprofloxacin versus 3 days 400 mg twice-daily of norfloxacin in uncomplicated urinary tract infection in women was designed. Patients underwent four visits: baseline, and at 3, 7 and 28 days. The main efficacy variables were clinical and microbiological outcome at day 7. Analyses on both valid patients and intention-to-treat populations were performed. Two hundred and twenty-six patients (114 receiving ciprofloxacin and 112 receiving norfloxacin) were considered valid for efficacy evaluation. Bacteriologic cure was 91.2% in the ciprofloxacin group and 91.9% in the norfloxacin group. Clinical resolution was 91.2 and 93.8%, respectively. Both treatments were equally efficacious (P = 0.016).


Proton transfer and proton concentrations in protonated Nafion fuel cell membranes.
Proton transfer in protonated Nafion fuel cell membranes is studied using several pyrene derivative photoacids. Proton transfer in the center of the Nafion nanoscopic water channels is probed with the highly charged photoacid 8-hydroxypyrene-1,3,6-trisulfonate (HPTS). At high hydration levels, both the time-integrated fluorescence spectrum and the fluorescence kinetics of HPTS permit the determination of hydronium concentration of the interior of the water pools in Nafion. The proton transfer kinetics of HPTS in protonated Nafion at maximum hydration are identical to the kinetics displayed by HPTS in a 0.5 M HCl solution. The hydronium concentration near the water interface in Nafion is estimated with rhodamine-6G to be 1.4 M. Excited state proton transfer (ESPT) is followed in the nonpolar side chain regions of Nafion with the photoacid 8-hydroxy-N,N,N',N',N'',N''-hexamethylpyrene-1,3,6-trisulfonamide (HPTA). Excited state proton transfer of HPTA is possible in protonated Nafion only at the highest hydration level due to a relatively high local pH.


Relationship between arterial stiffness and athletic training programs in young adult men.
We examined the relationships of endurance and strength exercise training and the adolescent duration of training to arterial stiffness in young adult men. We hypothesized that young adults participating in endurance sports would have decreased arterial stiffness, whereas those in strength-based sports would have increased arterial stiffness. In addition, we predicted that these trends would be more pronounced with an increase in the duration of sport participation. Subjects were male endurance-trained men with short (current age, 20 years; age at beginning of competitive sport, 15 years; sport careers, 5 years; n = 7, S-ET) and long (current age, 20 years; age at beginning of competitive sport, 12 years; sport careers, 8 years; n = 7, L-ET) competitive sport careers, strength-trained men with short (current age, 20 years; age at beginning of competitive sport, 16 years; sport careers, 4 years; n = 7, S-ST) and long (current age, 22 years; age at beginning of competitive sport, 15 years; sport careers, 7 years; n = 7, L-ST) careers, and sedentary control men (aged, 20 years; n = 7, C). The exercise training was associated with aortic pulse wave velocity (PWV), a traditional index of arterial stiffness, and the associations were statistically independent of blood pressure (BP). Aortic PWV was lower in L-ET than C and ST. Aortic PWV in L-ST was greater than that of C. The associations of exercise training with systemic arterial compliance (SAC), which inversely correlates with arterial stiffness, were also positive and BP independent. The SAC was greater in the ET groups compared with C and ST groups. The SAC in L-ST was lower than in C. These results suggest that changes in arterial stiffness associated with different training programs appear in young adults as well as in older humans, and these changes may begin in adolescence.


Treatment of open fractures with a computer-assisted external fixator system without the use of fluoroscopy.
Developed for the treatment of deformity correction, computer-assisted circular external fixators in recent years have also been used for fracture fixation. In this study, we aimed to present the efficacy of computer-assisted circular external fixator use in open long bone fractures with our new technique. Between February 2013 and June 2014, 14 patients (mean age 24.5 (range 20-32)) with open tibial or femoral open fractures were treated with the computer-assisted fixation system (Spider Frame-Tasarım Medikal, Istanbul, Turkey). In all patients, appropriate positions of the rings and Schanz screws were determined by measurements on preoperative radiographs. The length of the Schanz screws were determined by depth measure marks on drill bits. Obvious deformities were corrected intraoperatively by manipulations, but residual deformities were corrected by a software program (Spiderfix, Tasarım Medikal, Istanbul, Turkey). We did not use fluoroscopy during the procedures. Ten patients had tibia diaphyseal and four patients had femoral diaphyseal fractures. Mean surgical time was 24.2 (range 18-28) min. Average follow-up time was 10.2 (range 9-14) months. Mean time for deformity correction was 3.1 (2-5) days. Complete union was observed in all patients with a mean of 4.9 (range 3-9) months. There were two grade 2 pin site infections treated with oral antibiotherapy and pin site care. We did not detect any Schanz screw breakage, loosening, deep infection, nonunion, or malunion. Computer-assisted external fixation systems can be used in the treatment of open fractures, and they provide the chance for acute or gradual correction. Preoperative planning and assistant devices with depth measures may decrease the procedure time and the need for fluoroscopy use.


Pediatric Helicobacter pylori isolates display distinct gene coding capacities and virulence gene marker profiles.
Helicobacter pylori strains display remarkable genetic diversity, and the presence of strains bearing the toxigenic vacA s1 allele, a complete cag pathogenicity island (PAI), cagA alleles containing multiple EPIYA phosphorylation sites, and expressing the BabA adhesin correlates with development of gastroduodenal disease in adults. To better understand the genetic variability present among pediatric strains and its relationship to disease, we characterized H. pylori strains infecting 47 pediatric North American patients. Prevalence of mixed infection was assessed by random amplified polymorphic DNA analysis of multiple H. pylori clones from each patient. Microarray-based comparative genomic hybridization was used to examine the genomic content of the pediatric strains. The cagA and vacA alleles were further characterized by allele-specific PCR. A range of EPIYA motif configurations were observed for the cagA gene, which was present in strains from 22 patients (47%), but only 19 (41%) patients contained a complete cag PAI. Thirty patients (64%) were infected with a strain having the vacA s1 allele, and 28 patients (60%) had the babA gene. The presence of a functional cag PAI was correlated with ulcer disease (P = 0.0095). In spite of declining rates of H. pylori infection in North America, at least 11% of patients had mixed infection. Pediatric strains differ in their spectrum of strain-variable genes and percentage of absent genes in comparison to adult strains. Most children were infected with H. pylori strains lacking the cag PAI, but the presence of a complete cag PAI, in contrast to other virulence markers, was associated with more severe gastroduodenal disease.


Minocycline hyperpigmentation: skin, tooth, nail, and bone involvement.
The association of bone, tooth, nail and scleral pigmentation in a patient treated for acne with minocycline was noted. Hyperpigmentation appeared in old scar tissue, but not that present from recent surgical procedures.


Regulation of the 12-O-tetradecanoyl-phorbol-13-acetate-induced inhibition of intercellular communication.
Effects of the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and protein kinase C (PKC) inhibitors on cell-cell communication were studied in a normal rat liver cell line, clone 9. Communication was observed and quantitated with microspectofluorometric and image analysis techniques following scrape-loading of the cells with lucifer yellow. Lucifer yellow migrated as far as ten cells away from the scraped edge in control populations. Two minute TPA (25-50 micrograms/ml) treatment inhibited dye movement such that the dye remained mainly in the cells at the cut edge. The TPA-induced inhibition of cell-cell communication could be partially blocked by 15 min treatment of the cell populations with the PKC inhibitors trifluoperazine (30 micrograms/ml), staurosporine (2 x 10(-8) or 2 x 10(-6) M), sangivamycin (15 or 200 microM), or a PKC inhibitor peptide (20 micrograms/ml) scraped in at the same time as lucifer yellow. Normal communication was observed in cultures treated only with PKC inhibitors. Lower concentrations of TPA (50 ng/ml-1 micrograms/ml) used for 2 min did not inhibit dye communication. Our results demonstrate the phorbol ester-induced interruption of cell-cell communication. The inhibition of PKC by inhibitors eliminates the effect of TPA on communication. Our data are consistent with a role of PKC in the control of junctional communication.


Development of new hydroactive dressings based on chitosan membranes: characterization and in vivo behavior.
Different poly(vinyl alcohol) (PVA)/chitosan lactate (ChL)-blended hydrogels containing nitrofurazone as a local anti-infective drug were prepared by the phase-inversion technique. The swelling degree, surface free energy, mechanical properties, and nitrofurazone release of these membranes were determined. Blood compatibility of these systems was evaluated by the open-static platelet adhesion test with whole human blood. The results showed that water absorption into the PVA/ChL membranes slowed down, governed by the rate at which the dressing interacted with the physiological fluid. Swelling degree values up to 200% were observed. The rate of release of nitrofurazone seemed to depend on the ChL percentage on the blend as well as the pH of the solution. The surface free energy values were in the range of 20-30 dynes/cm, which was appropriate for a favorable interaction with blood. From the Young's module curve, it could be seen that elastic hydrogels were obtained with increment of ChL in the PVA/ChL blends. Values of platelet adhesion and whole blood clotting times for the PVA/ChL blends as well as the increase of ChL, which appears to reduce the fibrinogen adsorption on the PVA/ChL membranes, demonstrated that the blood compatibility of PVP/ChL blends is superior to that separated polymers. The results of in vivo experiments in rats were in very good agreement with these observations, suggesting that PVA/ChL may serve as a new type of potential wound-dressing material.


Serum albumin levels and their correlates among individuals with motor disorders at five institutions in Japan.
The level of serum albumin is an index of nourishment care and management. However, the distribution and correlates of serum albumin levels among individuals with motor disorders have not been reported until now. Therefore, we examined the distribution and correlates of serum albumin levels among individuals with motor disorders. A cross-sectional study on 249 individuals with motor disabilities (144 men, mean age: 51.4 years; 105 women, mean age: 51.4 years) was conducted at five institutions in Ibaraki Prefecture, Japan in 2008. The results were compared with data from the National Health and Nutrition Survey. The mean serum albumin levels were 4.0 ± 0.4 g/dL for men and 3.8 ± 0.5 g/dL for women. Overall, 17 (11.8%) men and 25 (23.8%) women had hypoalbuminemia (serum albumin level ≤ 3.5 g/dL); these proportions were greater than those among healthy Japanese adults (≤ 1%). Low serum albumin level was related with female sex, older age, low calf circumference, low relative daily energy intake, low hemoglobin (Hb), low blood platelet count, low high-density lipoprotein cholesterol (HDL-C), low HbA1c, and high C-reactive protein (CRP) levels. The strongest correlates, based on standardized betas, were Hb (0.321), CRP (-0.279), and HDL-C (0.279) levels. These results indicate that the prevalence of hypoalbuminemia is higher in individuals with motor disabilities than in healthy individuals and that inflammation is a strong negative correlate of serum albumin levels. Therefore, inflammation should be examined for the assessment of hypoalbuminemia among institutionalized individuals with motor disabilities.


High dose glycine nutrition affects glial cell morphology in rat hippocampus and cerebellum.
Enhancement of N-methyl-D-aspartate (NMDA) receptor-mediated neurotransmission by glycine (Gly) administration may represent a novel pharmacological strategy in schizophrenia. Given the involvement of NMDA receptors in plasticity and excitatory processes, the present study explores effects of Gly on brain cell morphology. Adult rats were randomized to receive, for 2 wk, no dietary supplementation or supplementation with 0.8 or 3.2 g/kg per day Gly. Glial cell morphology was examined using antibodies to glial fibrillary acidic protein (GFAP) and to microglial complement receptor 3 (CR3). Cresyl violet was used for general cellular staining. No evidence of neuronal or microglial pathology was found. Although astrocyte proliferation was not evident in Gly-treated rats, GFAP-like immunoreactivity was dose-dependently increased in the hippocampus (p<0.01), whereas in cerebellum, a dose dependent decrease in density of astocytic fibres was demonstrated (p<0.01). These findings demonstrate for the first time that in vivo administration of high dose Gly may induce brain morphology changes.


Hybrid incompatibilities in the parasitic wasp genus Nasonia: negative effects of hemizygosity and the identification of transmission ratio distortion loci.
The occurrence of hybrid incompatibilities forms an important stage during the evolution of reproductive isolation. In early stages of speciation, males and females often respond differently to hybridization. Haldane's rule states that the heterogametic sex suffers more from hybridization than the homogametic sex. Although haplodiploid reproduction (haploid males, diploid females) does not involve sex chromosomes, sex-specific incompatibilities are predicted to be prevalent in haplodiploid species. Here, we evaluate the effect of sex/ploidy level on hybrid incompatibilities and locate genomic regions that cause increased mortality rates in hybrid males of the haplodiploid wasps Nasonia vitripennis and Nasonia longicornis. Our data show that diploid F(1) hybrid females suffer less from hybridization than haploid F(2) hybrid males. The latter not only suffer from an increased mortality rate, but also from behavioural and spermatogenic sterility. Genetic mapping in recombinant F(2) male hybrids revealed that the observed hybrid mortality is most likely due to a disruption of cytonuclear interactions. As these sex-specific hybrid incompatibilities follow predictions based on Haldane's rule, our data accentuate the need to broaden the view of Haldane's rule to include species with haplodiploid sex determination, consistent with Haldane's original definition.


Pectin methylesterase of Datura species, purification, and characterization from Datura stramonium and its application.
Pectin methylesterases (PME; EC 188.8.131.52) involved in de-esterification of pectin and have applicability in food, textiles, wines, pulp, and paper industries. In the present study, we compared PME activity of different parts of 3 Datura species and found that fruit coat showed maximum PME activity followed by leaf and seed. PME from leaves of D. stramonium (DsPME) was purified and characterized. DsPME showed optimum activity at 60 °C and pH 9 in the presence of 0.3 M NaCl. DsPME was stable at 70 °C and retained more than 40% activity after 60 min of incubation. However, enzyme activity completely abolished at 80 after 5 min of incubation. It follows Michaelis-Menten enzyme kinetics. Km and Vmax with citrus pectin were 0.008 mg/ml and 16.96 µmol/min, respectively. DsPME in combination with polygalactourenase (PGA) increased the clarity of orange, apple, pomegranate and pineapple juices by 2.9, 2.6, 2.3, and 3.6 fold, respectively in comparison to PGA alone. Due to very high de-esterification activity, easy denaturation and significant efficacy in incrementing clarification of fruit juice makes DsPME useful for industrial application.


Rechargeable Room-Temperature Na-CO2 Batteries.
Developing rechargeable Na-CO2 batteries is significant for energy conversion and utilization of CO2 . However, the reported batteries in pure CO2 atmosphere are non-rechargeable with limited discharge capacity of 200 mAh g(-1) . Herein, we realized the rechargeability of a Na-CO2 battery, with the proposed and demonstrated reversible reaction of 3 CO2 +4 Na↔2 Na2 CO3 +C. The battery consists of a Na anode, an ether-based electrolyte, and a designed cathode with electrolyte-treated multi-wall carbon nanotubes, and shows reversible capacity of 60000 mAh g(-1) at 1 A g(-1) (≈1000 Wh kg(-1) ) and runs for 200 cycles with controlled capacity of 2000 mAh g(-1) at charge voltage <3.7 V. The porous structure, high electro-conductivity, and good wettability of electrolyte to cathode lead to reduced electrochemical polarization of the battery and further result in high performance. Our work provides an alternative approach towards clean recycling and utilization of CO2 .


The trypanosome-specific proteins FPRC and CIF4 regulate cytokinesis initiation by recruiting CIF1 to the cytokinesis initiation site.
The evolutionarily early divergent human parasite Trypanosoma brucei proliferates through binary cell fission in both its tsetse fly vector and mammalian host. The parasite divides unidirectionally along the longitudinal cell axis from the anterior cell tip toward the posterior cell tip through a mechanism distinct from that in the cells of its human host. Initiation of cytokinesis in T. brucei is regulated by two evolutionarily conserved protein kinases, the Polo-like kinase TbPLK and the Aurora B kinase TbAUK1, and a cohort of trypanosome-specific proteins, including the three cytokinesis initiation factors CIF1, CIF2, and CIF3. Here, using RNAi, in situ epitope tagging of proteins, GST pulldown, and coimmunoprecipitation assays, and immunofluorescence and scanning electron microscopy analyses, we report the identification and functional characterization of two trypanosome-specific proteins, flagellum attachment zone tip-localizing protein required for cytokinesis (FPRC) and CIF4. We found that the two proteins colocalize to the distal tips of the new and the old flagellum attachment zones and are required for cytokinesis initiation. Knockdown of FPRC or CIF4 disrupted the localization of CIF1, suggesting that they function upstream of CIF1. Moreover, depletion of CIF4 abolished FPRC localization, indicating that CIF4 acts upstream of FPRC. Together, these results identify two new cytokinesis regulators in T. brucei and integrate them into the CIF1-mediated cytokinesis regulatory pathway. These findings highlight the existence of a cytokinesis pathway in T. brucei that is different from that of its mammalian host and therefore suggest that cytokinesis in T. brucei could potentially be exploited as a new drug target.


[Synovial membrane diagnostic assessment in rheumatoid arthritis].
Rheumatoid arthritis (RA) is the most frequent inflammatory rheumatic disease. At the beginning of the disease, where, based on today's knowledge the therapeutic possibilities are largest, the diagnostic methods do not permit a differentiated estimation of the prognosis. Conventional x-rays are mostly normal and serum markers unspecific. So far--in contrast to other diseases--only little information had been drawn from the pathomorphologic substrate "synovialis" itself to assess the prognosis. Reasons therefor were found in difficulties in obtaining synovial tissue besides surgical interventions, particularly in patients with early arthritis. By minimalizing the diagnostic instruments and improvement of the technique, synovial tissue sampling in RA has become minimally invasive and it is even possible to perform on the smallest joints, such as finger joints. Hereby, synovial analysis is open for detecting pathways of inflammation and joint destruction, which might support the advancement of new therapeutic strategies, followed by a better prognosis and outcome of RA.


Formative research for the development of an Eye Health Strategic Planning and Evaluation Framework and a Checklist: A health systems approach.
This formative research process aimed to develop an Eye Health Strategic Planning and Evaluation Framework and indicator Checklist. The research process utilized a multi-phased multiple methods approach including literature review, initial expert review (n = 27), findings from a Cambodian Avoidable Blindness Initiative demonstration project (2009-2012), observation and analysis of four rural sites of the Indian LV Prasad Eye Institute Pyramid Model (n = 21), and finally, a critique by Cambodian government eye health professionals/staff (n = 15), health center staff and community representatives (n = 77) and patients (n = 62). Results from three Cambodian population-based surveys (KAP n = 599, patient follow-up n = 354, and RAAB 4650) also informed the development of the Framework and the Checklist. The Framework domains include: situation analysis, determinants of accessibility, service delivery systems, operation systems, networks and linkages, outcomes, and impact. Domains were subdivided into 59 components. The Checklist consists of 253 indicator items. The Eye Health Strategic Planning and Evaluation Framework and the Checklist can assist policy makers, program planners, and evaluators to develop a comprehensive whole of systems approach to eye health care to improve coverage and utilization of services.


Synthesis and characterization of xanthene-bridged Schiff-base dimanganese(III) complexes: bimetallic catalysts for asymmetric oxidation of sulfides.
Dimanganese(III) complexes of salen-type ligands anchored by 9,9-dimethylxanthene-4,5-diyl spacers were synthesized. Two types of structures, cyclic and acyclic forms, are presented. The 2 + 2 Schiff-base condensation of 5,5'-(9,9-dimethylxanthene-4,5-diyl)bis(salicylaldehyde) and 1,2-diaminobenzene gave a macrocyclic ligand, from which a cyclic dimanganese(III) complex was synthesized. A similar dimanganese(III) complex with stereogenic centers was synthesized by the metal-assisted condensation reaction using (1R,2R)-1,2-diaminocyclohexane. Acyclic dimanganese(III) complexes anchored by one xanthene spacer were obtained by metal-assisted stepwise condensation reactions using the xanthene-bridged bis(salicylaldehyde), (1R,2R)-1,2-diaminocyclohexane monohydrochloride, and salicylaldehydes. X-Ray crystal structure analysis confirmed that both cyclic and acyclic complexes have two cofacially oriented salen-type manganese(III) units in a complex cation, showing an Mn...Mn distance of ca. 5.1 A. Asymmetric oxidation of sulfides was performed by using the chiral dimeric complexes as catalysts, which showed enantiomeric excesses ranging from 5 to 19%. The addition of 4-(dimethylamino)pyridine to the reaction system improved the enantioselectivity up to 39% ee. This effect was not observed for the corresponding mononuclear catalyst.


In Vitro Activity and MIC of Sitafloxacin against Multidrug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis Isolated in Thailand.
New fluoroquinolones (FQs) have been shown to be more active against drug-resistant Mycobacterium tuberculosis strains than early FQs, such as ofloxacin. Sitafloxacin (STFX) is a new fluoroquinolone with in vitro activity against a broad range of bacteria, including M. tuberculosis This study aimed to determine the in vitro activity of STFX against all groups of drug-resistant strains, including multidrug-resistant M. tuberculosis (MDR M. tuberculosis), MDR M. tuberculosis with quinolone resistance (pre-XDR), and extensively drug-resistant (XDR) strains. A total of 374 drug-resistant M. tuberculosis strains were tested for drug susceptibility by the conventional proportion method, and 95 strains were randomly submitted for MIC determination using the microplate alamarBlue assay (MABA). The results revealed that all the drug-resistant strains were susceptible to STFX at a critical concentration of 2 μg/ml. Determination of the MIC90s of the strains showed different MIC levels; MDR M. tuberculosis strains had a MIC90 of 0.0625 μg/ml, whereas pre-XDR and XDR M. tuberculosis strains had identical MIC90s of 0.5 μg/ml. Common mutations within the quinolone resistance-determining region (QRDR) of gyrA and/or gyrB did not confer resistance to STFX, except that double mutations of GyrA at Ala90Val and Asp94Ala were found in strains with a MIC of 1.0 μg/ml. The results indicated that STFX had potent in vitro activity against all the groups of drug-resistant M. tuberculosis strains and should be considered a new repurposed drug for treatment of multidrug-resistant and extensively drug-resistant TB.


Changes in myocardial electrical impedance induced by coronary artery occlusion in pigs with and without preconditioning: correlation with local ST-segment potential and ventricular arrhythmias.
Myocardial ischemia increases tissue electrical resistivity leading to cell-to-cell uncoupling, and this effect is delayed by ischemic preconditioning in isolated myocardium. Alterations in myocardial resistivity elicited by ischemia in vivo may influence arrhythmogenesis and local ST-segment changes, but this is not well known. Myocardial impedance (resistivity [omega x cm] and phase angle [degrees]), epicardial ST segment, and ventricular arrhythmias were analyzed during 4 hours of coronary artery occlusion in 11 anesthetized open-chest pigs; these were compared with 13 other pigs submitted to a similar coronary occlusion preceded by ischemic preconditioning. Myocardial resistivity rose slowly during the first 34+/-7 minutes of occlusion (237+/-41 to 359+/-59 omega x cm), increased rapidly to 488+/-100 omega x cm at 60 minutes, and reached a plateau value (718+/-266 omega x cm, ANOVA; P<.01) at 150+/-69 minutes. By contrast, phase-angle changes began after 17 minutes of ischemia (-3.0+/-1.6 degrees to -4.2+/-1.2 degrees at 29+/-8 minutes) and evolved faster thereafter (-12.5+/-5.3 degrees at 144+/-56 minutes). Marked changes in myocardial impedance were observed during the reversion of ST-segment elevation that occurred 1 to 4 hours after occlusion, but impedance changes were less apparent during the early ST-segment recovery seen at 15 to 35 minutes of ischemia. The second arrhythmia peak (30+/-5 minutes) coincided with the fast change in tissue impedance, and both were delayed (P<.05) by ischemic preconditioning. A rapid impairment of myocardial impedance occurs after 30 minutes of coronary occlusion, and its onset is better defined by shift in phase angle than by rise in tissue resistivity. Phase 1b arrhythmias are associated with marked impedance changes, and both are delayed by preconditioning. Reversion of ST-segment elevation is partially associated with impairment of myocardial impedance, but other factors play a role as well.


Baroreflex activation therapy in patients with pre-existing implantable cardioverter-defibrillator: compatible, complementary therapies.
The Neo™ System (CVRx) is an implantable device, CE certified for the treatment of resistant hypertension and investigationally used to treat systolic heart failure by electrical stimulation of the carotid baroreceptors. It is unknown whether interaction might exist between the Neo System and implantable cardioverter-defibrillators (ICDs). Compatibility of the Neo device was tested in seven consecutive patients with pre-existing ICDs. Intra- and post-operative testing was completed with ICD and Neo settings programmed to provoke interaction. Intracardiac electrograms were printed to determine interaction with the ICD. Interaction testing during implantation and follow-up showed that there was no device-device interaction. No interaction was observed at maximum atrial and ventricular sensitivity settings and maximum Neo output settings. Combined therapy with the Neo device and at least in this study reported that transvenous ICD systems can be performed safely.


Aluminum, a Friend or Foe of Higher Plants in Acid Soils.
Aluminum (Al) is the most abundant metal in the earth's crust, but its availability depends on soil pH. Despite this abundance, Al is not considered an essential element and so far no experimental evidence has been put forward for a biological role. In plants and other organisms, Al can have a beneficial or toxic effect, depending on factors such as, metal concentration, the chemical form of Al, growth conditions and plant species. Here we review recent advances in the study of Al in plants at physiological, biochemical and molecular levels, focusing mainly on the beneficial effect of Al in plants (stimulation of root growth, increased nutrient uptake, the increase in enzyme activity, and others). In addition, we discuss the possible mechanisms involved in improving the growth of plants cultivated in soils with acid pH, as well as mechanisms of tolerance to the toxic effect of Al.


Molecular diagnosis of visceral leishmaniasis: a French study comparing a reference PCR method targeting kinetoplast DNA and a commercial kit targeting ribosomal DNA.
PCR revolutionized the direct diagnosis of infectious diseases, especially protozooses, where the infectious load is usually low. Commercial PCR methods are available and offer many advantages, including convenience and batch tracking as part of a quality system. For most parameters, the performance of commercial methods is at least as good as that of finely optimized methods developed in expert laboratories. This comparison work has not been done for the molecular diagnosis of visceral leishmaniasis. Leishmania sp. has a unique organelle, the kinetoplast, which corresponds to the mitochondrial DNA. It is organized into a large number of minicircles, which has made it a target for the development of diagnostic PCR. The quanty Leishmaniae, Clonit kit targeting ribosomal DNA was compared to a widely used laboratory-developed method based on kinetoplast DNA. This reference method gave significantly better results, probably due to the difference in the number of repeats of the PCR targets.


Determining the site of origin of mucinous adenocarcinoma: an immunohistochemical study of 175 cases.
Mucinous adenocarcinomas (MAs) of various origins may have a similar histologic appearance and frequently metastasize to distant sites, which often causes diagnostic problems in surgical pathology practice. The immunohistochemical profiles of MAs of various origins have not been well studied. We investigated the expression of 10 immunohistochemical markers (CK7, CK20, CDX-2, β-catenin, MUC-1, MUC-2, MUC-6, ER, WT-1, and PAX-8) in 175 cases of MA, including 69 cases from the lower gastrointestinal (GI) tract, 41 from the upper GI tract, 27 from gynecologic organs, 4 from the urinary bladder, 18 from the breast, and 16 from the lung. We found that lower GI MAs (colon, rectum, and anus) frequently expressed CDX-2 (42 of 42, 100%; 33 of 42 with homogenous positivity, 79%), MUC-2 (42 of 42; 100%), CK20 (41 of 42; 98%), and β-catenin (nuclear) (27 of 42; 64%) and rarely expressed MUC-6 (2 of 42; 5%) and CK7 (8 of 42; 19%). Most of the CK7-positive cases were from the rectum and anus (7 of 8; 88%). The expression of these markers in appendiceal MAs was similar to that of low GI tract MAs, except for a lower percentage of homogenous CDX-2 (3 of 27; 11%) and nuclear β-catenin (3 of 27; 11%) expression. Unlike their lower GI tract counterparts, the upper GI tract MAs (ampulla, pancreas/biliary tree, and stomach/esophagus) frequently expressed CK7 (38 of 41; 93%) and MUC-6 (31 of 41; 76%) and were rarely homogenously positive for CDX-2 (4 of 41; 10%) and nuclear positive for β-catenin (8 of 41; 19%). Breast MAs were frequently positive for CK7 (18 of 18; 100%), MUC-1 (18 of 18; 100%), MUC-2 (18 of 18; 100%), ER (16 of 18; 89%), MUC-6 (9 of 18; 50%), and WT-1 (9 of 18; 50%). Lung MAs were frequently positive for CK7 (16 of 16; 100%) and MUC-1 (15 of 16; 94%). Gynecologic MAs were positive for CK7 (25 of 27; 93%) and PAX-8 (13 of 27; 48%). We conclude that homogenous CDX-2 and nuclear β-catenin expressions are commonly seen in lower GI tract MAs. In contrast, appendiceal MAs are usually heterogenously positive for CDX-2 and show cytoplasmic positivity for β-catenin. Unlike lower GI tract MAs, upper GI tract MAs are frequently positive for CK7 and MUC-6. As is the case in appendiceal MAs, the upper GI tract MAs may also be heterogenously positive for CDX-2. Breast MAs are positive for ER and WT-1, whereas gynecologic MAs are positive for PAX-8 and negative for WT-1.


Pseudocirrhosis in Breast Cancer - Experience From an Academic Cancer Center.
Pseudocirrhosis is characterized by radiological changes in the liver that resemble cirrhosis, but with more rapid onset and progression. Though reported most frequently in patients with metastatic breast cancer, little is known about its prognostic factors and impact on breast cancer outcomes. In this observational study, we reviewed abdominal CT and/or MRI scan reports of all patients with invasive breast cancer diagnosed at our center, during a ten-year period, to identify patients with pseudocirrhosis. Exclusion criteria included lack of baseline imaging, pre-existing cirrhosis, hepatitis B or C, other chronic liver diseases, or heavy alcohol use. Routine descriptive statistical measures were used. Survival distributions were estimated using Kaplan-Meier method, and Cox regression was used for multivariate analysis. Two-tailed p < 0.05 was considered significant. Eighty-six patients were included - all were females, median age was 57.5 years, and 90% were Caucasian; 86% of primary tumors were hormone-receptor positive and 17% were HER2 positive. Most patients (98%) had metastatic disease with liver involvement (94%), and were heavily pre-treated - 97% with chemotherapy, 85% with hormonal therapy, and 19% with anti-HER2 agents. Median interval from breast cancer diagnosis to pseudocirrhosis was 75.4 months (IQR 35.2-115.3 months). Thirty-six percentage of patients had ≥1 signs of portal hypertension and 49% had ≥1 signs of hepatocellular failure. Pseudocirrhosis led to permanent discontinuation of chemotherapy, endocrine therapy, and all systemic therapies in 29%, 31%, and 20% patients, respectively. Median overall survival from diagnosis of pseudocirrhosis was 10.0 months (95%CI 5.2-14.8 months). On multivariate analysis, coagulopathy, hyperbilirubinemia, hypoalbuminemia, and cancer progression were independently predictive of mortality. In this largest series, to date, of breast cancer with pseudocirrhosis, the latter was often complicated by portal hypertension and hepatocellular failure, and markedly impacted breast cancer management. Survival was shorter for patients who developed hepatocellular failure.


Circulating levels of soluble interleukin-6 receptor in patients with bronchial asthma.
In the search for markers of airway inflammation, we investigated the role of soluble interleukin-6 receptor (sIL-6R) in patients with bronchial asthma. Serum levels of sIL-6R were measured in 20 patients with stable asthma and in 18 healthy control subjects by means of a sandwich enzyme-linked immunosorbent assay. Such levels were also evaluated during a spontaneous attack of asthma (n = 10) as well as that after allergen inhalation (n = 7). Results were compared with those observed during the stable state and after the inhalation of methacholine. Serum levels of sIL-6R in asthmatic patients (132 +/- 31 ng/ml) significantly exceeded those of control subjects (111 +/- 16 ng/ml) (p < 0.05). These levels showed no correlation with such clinical variables as nonspecific bronchial hyperreactivity, atopic status, or serum concentration of IgE. Serum sIL-6R levels observed during an asthmatic attack versus those during the stable state (4 wk later) differed significantly. After a severe attack of asthma, such levels were significantly elevated on the second and third days, but not on Day 5. After challenge, circulating levels of sIL-6R were significantly increased 24 h after the inhalation of allergen but not of methacholine. Results suggest that serum levels of sIL-6R are increased in patients with asthma and are further increased during a spontaneous attack or that provoked by the inhalation of allergen. Thus, serum sIL-6R may reflect inflammation of the airway. Further studies are indicated to determine the clinical significance and the application of serum levels of sIL-6R in evaluating asthmatic patients.


Loxosceles intermedia spider envenomation induces activation of an endogenous metalloproteinase, resulting in cleavage of glycophorins from the erythrocyte surface and facilitating complement-mediated lysis.
Loxosceles is the most venomous spider in Brazil, and envenomation causes dermonecrosis and complement (C)-dependent intravascular hemolysis. The authors studied the mechanism of induction of C-induced hemolysis. Purified Loxosceles toxins rendered human erythrocytes susceptible to lysis by human C but did not have an effect on the E-bound C-regulators DAF, CR1, or CD59. However, incubation with venom toxins caused cleavage of glycophorin from the erythrocyte (E) surface, facilitating C activation and hemolysis. The results suggest that glycophorin is an important factor in the protection of E against homologous C. Cleavage of glycophorin (GP) A, GPB, and GPC occurred at sites close to the membrane but could not be accomplished using purified GPA and purified toxins, demonstrating that cleavage was not an effect of a direct proteolytic action of the Loxosceles toxins on the glycophorins. Inhibition of the cleavage of glycophorins induced by Loxosceles venom was achieved with 1,10-phenanthroline. The authors propose that the sphingomyelinase activity of the toxins induces activation of an endogenous metalloproteinase, which then cleaves glycophorins. They observed the transfer of C-dependent hemolysis to other cells, suggesting that the Loxosceles toxins can act on multiple cells. This observation can explain the extent of hemolysis observed in patients after envenomation. Identification of the mechanism of induction of susceptibility to C-mediated lysis after Loxosceles envenomation opens up the possibility of the development of an effective therapeutic strategy. (Blood. 2000;95:683-691)


Interleukin 1 induces an autocrine loop hepatocyte growth factor/c-Met in murine Kaposi-like spindle cells.
Several cytokines, growth factors and the HIV transactivator Tat were shown to be involved in the pathogenesis of Kaposi's sarcoma. BKV/tat transgenic mice develop Kaposi's sarcoma-like lesions, and spindle-shaped cells (TTB) have been derived from these lesions. Here we show that TTB cells co-express hepatocyte growth factor (HGF) and its receptor, the product of the oncogene c-Met. An autocrine loop HGF/Met sustains spindle cell proliferation in vitro; indeed, an antisense oligomer targeted against HGF markedly inhibited cell growth. Moreover, HGF and Met are overexpressed after exposing TTB cells to the proinflammatory cytokine interleukin 1 (IL-1). We argue that upon exposure to IL-1, an HGF/Met autocrine loop is induced which could explain the appearance of multiple foci of uncontrolled growth. In addition, due to its angiogenic activity, HGF may also sustain the neovascularization typical of Kaposi's sarcoma lesions.


Inhomogeneity of gated and ungated SPECT technetium-99m-sestamibi bull's-eyes in normal dogs: comparison with thallium-201.
By computer simulation, we have previously hypothesized, independent of the isotope imaged, that differences in view-to-view resolution and attenuation patterns predictably cause count density distortions in SPECT images. We tested the simulation predictions for both ECG-gated and ungated SPECT 99mTc-sestamibi and SPECT 201Tl myocardial perfusion images in normal dogs. In agreement with the predictions of the computer model, distortions in SPECT 99mTc-sestamibi myocardial perfusion images are virtually equivalent to SPECT 201Tl, dependent on the exact SPECT acquisition orbit and markedly different for a posterior 180 degrees acquisition arc compared to an anterior 180 degrees acquisition arc. Furthermore, ungated and gated SPECT 99mTc-sestamibi images show similar count inhomogeneities. These results suggest that little is to be gained from a 360 degrees acquisition with SPECT 99mTc-sestamibi, and that image distortions from gated or ungated SPECT 99mTc images with 180 degrees orbits will be similar to those in SPECT 201Tl images.


The need to differentiate between adults and children when treating severe asthma.
Severe asthma at all ages is heterogeneous incorporating several phenotypes that are distinct in children and adults, however, there are also numerous similar features including the limitation that they may not remain stable longitudinally. Severe asthma in both children and adults is characterized by eosinophilic airway inflammation and evidence of airway remodeling. In adults, targeting eosinophilia with anti-IL-5 antibody therapy is very successful, resulting in the recommendation that sputum eosinophils should be used to guide treatment. In contrast, data for the efficacy of blocking IL-5 remain unavailable in children. However, its effectiveness is uncertain since many children with severe asthma have normal blood eosinophils and the dominance of Th2-mediated inflammation is controversial. Approaches that have revealed gene signatures and biomarkers such as periostin that are specific to adult disease now need to be adopted in children to identify effective pediatric specific therapeutics and minimize the extrapolation of adult therapeutics to children.


Resisting Aridification: Adaptation of Sap Conduction Performance in Moroccan Wild Olive Subspecies Distributed Over an Aridity Gradient.
In the current context of global change, the increasing frequency and the length of drought periods are testing the resistance capacities of plants of dry habitats. However, although the adaptation of plants to drought has been widely studied, the anatomical features of wood influencing the functional responses of plants to drought are still lacking at the intraspecific level, especially for species with a wide geographical distribution. As a result, we have studied the variation of wood anatomical traits related to sap conduction (i.e., vessel surface area, vessel density, and number of vessels joined by radial file) in two wild olive subspecies distributed in Morocco (i.e., Olea europaea subsp. europaea. var. sylvestris and Olea europaea subsp. maroccana), in relation to various drought conditions. This functional study, based on wood trait measurements of 351 samples from 130 trees and 13 populations, explores potential sap conduction in relation to environmental parameters and as a result, strategies to resist water stress. We found that (1) branch diameter (BD) captured 78% of total wood trait variation, (2) vessel size (SVS) expressed 32% of intraspecific variation according to cambium age, and (3) the positive relationship between SVS and BD could be explained by climate type, vegetation cover changes, and therefore available water resources. Taking into consideration the diameter of the branch as the main factor of anatomical variation, established reaction norms (linear models) at the intrapopulation scale of vessel lumen area according to aridity show for the first time how the functioning of the cambium modulates and controls sap conduction, according to aridity and thus available water resources. They pinpoint the risks incurred by the wild olive tree in the perspective of a dramatic increase in aridity, in particular, the inability of the cambium to produce large enough vessels to efficiently transport sap and irrigate the leaves. Finally, this study opens new and interesting avenues for studying at a Mediterranean scale, the resistance and the vulnerability of wild forms and cultivated varieties of olive to heterogeneous and changing environmental conditions.


Tagraxofusp followed by combined azacitidine and venetoclax in blastic plasmacytoid dendritic cell neoplasm: A case report and literature review.
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and aggressive hematologic malignancy that originates from plasmacytoid dendritic cells. It can involve skin, bone marrow, and/or lymph nodes. There is no consensus recommendation regarding treatment especially in the relapsed setting. Tagraxofusp, a CD123 directed agent, was recently approved by the Food and Drug Administration to treat BPDCN. We report a case of an elderly patient with diagnosis of BPDCN who was treated initially with tagraxofusp followed by azacitidine and venetoclax combination on relapse. A 79 year old male presented with violaceous skin lesions. He had no other symptoms. Biopsy of these lesions was consistent with a diagnosis of BPDCN. Further testing showed no extracutaneous involvement.Management and outcome: Tagraxofusp was started at full dose (12 mcg/kg). This dose was not tolerated well. Patient could only tolerate the lowest dose (5 mcg/kg). Toxicities included elevated liver function tests, hyperglycemia, capillary leak syndrome, and pancreatitis. Dose escalation on progression was not possible due to side effects. Treatment was switched to venetoclax and azacitidine. Combination treatment was tolerated very well and patient showed major cutaneous response after 5 cycles and continues to do well. Tagraxofusp is a novel therapy that needs more real-world experience. This case describes the clinical course of an elderly patient on tagraxofusp. We also review the literature of azacytidine/venetoclax combination as a potential yet tolerable treatment option for this rare disease entity. This is the fourth case in literature to be treated with this combination.


Inhibitory effects of chitosan coating combined with organic acids on Listeria monocytogenes in refrigerated ready-to-eat shrimps.
Chitosan and organic acids are natural preservatives with proven antimicrobial efficacies. This study investigated the effects of chitosan coating combined with organic acids on inhibiting Listeria monocytogenes in ready-to-eat (RTE) shrimps during storage at 4°C. Cooked shrimps inoculated with L. monocytogenes were coated with 0.5 and 1 % chitosan solutions in combination with 2 % organic acids (acetic, citric, lactic, and malic acids) for 5 min and then stored at 4°C for 16 days. Bacterial counts, pH, and sensory properties were analyzed every 4 days. The results indicated that the antimicrobial effects of chitosan were significantly enhanced (P < 0.05) when it was combined with 2 % acetic, citric, lactic, or malic acid. Chitosan at 1 % combined with 2 % acetic acid was the most effective treatment, which caused a 5.38-log CFU/g bacterial reduction after 16 days in comparison with the controls. Transmission electron microscopy further confirmed the enhanced antimicrobial effects of the combination of chitosan and organic acids. Such combined treatments also maintained the sensory properties of RTE shrimps. The use of chitosan coating combined with organic acids significantly lowered (P < 0.05) the pH values of RTE shrimps compared with the use of chitosan alone. Therefore, we conclude that chitosan coating combined with acetic acid could be a promising antimicrobial method to prevent the proliferation of L. monocytogenes in RTE shrimps with extended shelf life.


[Increased promoter activity of human delta-globin gene with point mutation of C-T at site-64].
With PCR point mutation technique, the CCAAC box in the promoter region of delta-globin gene was changed into CCAAT which is required in the normal transcription of beta-globin gene at the corresponding site. The normal and mutant segments of the delta-globin promoter region obtained with each segment of about 300 bp (-280/+54 bp), a fragment sufficient for the requirement of its normal promoter function, were separately cloned into pUC19/Hinc II. After sequencing, it showed that the CCAAC box had been successfully changed into CCAAT without other mutations. The two segments were released by Nco I and BamH I, and were cloned into the corresponding site of plasmid pMG3 which has the firefly luciferase as its reporter gene. The positive clones were selected by Dot Blot and were comfirmed with restriction mapping. Then the two recombinant plasmids were transfected into HeLa cell line respectively and the promoter activities of the two fragments were identified by comparing the light intensity generated by the cell extracts when added with luciferase assay substrate. The results demonstrated that the luminescence of the mutant group is on average five times higher than that of the normal group, which indicate that the C-T mutation of the CCAAC box of delta-globin promoter region could increase its promoter activity. Our results have confirmed the hypothesis that the sequence of CCAAC box of human delta-globin gene is one of the main reasons which accounts for its low expression level. The author is now studying the specific DNA binding protein of the upstream promoter region of human delta-globin gene.


Complex regional pain syndrome: the role of the psychiatrist as an expert witness.
Complex regional pain syndrome (CRPS) is a regional pain syndrome of unclear pathophysiology, typically affecting the hand or foot. CRPS can be subject to personal injury litigation and may be the cause of substantial future loss of earnings claims. In clinical practice, a variety of medical experts can be involved in the diagnosis and management of CRPS and this can lead to confusion about the most appropriate expert for lawyers to instruct. This paper defines the role of psychiatrists as experts to the court in CRPS and reviews the associated psychiatric literature. Psychiatrists do not have expertise in the clinical diagnosis of CRPS, but they are able to assess the presence of co-morbid psychiatric conditions. They can assist the court as experts in the underlying neuro-physiological and psychological processes which may explain some of the important features of CRPS and an individual's psychological vulnerability to CRPS. Their expertise in psychological and psycho-pharmacological interventions should be considered for any multidisciplinary treatment programme for CRPS along with the likely contribution of these interventions to the overall prognosis.


A possible extratubular origin of epididymal basal cells in mice.
The origin of basal cells in mouse epididymis was examined by counting the numbers of basal cells, intratubular mitotic figures and peritubular cells during development of the epididymis. Putative precursors of basal cells were labelled with bromodeoxyuridine and the nuclei of daughter cells were examined. Histochemical localization of cytokines was performed to gauge their involvement in migration of basal cell precursors from extratubular sources. The results indicate that basal cells may arise from extratubular sources as: (i) there was a decrease in the number of mitotic figures as the number of basal cells increased; (ii) no mitotic figures were observed in the base of the epithelium; (iii) the increase in the number of peritubular cells did not parallel the number of basal cells in all epididymal regions; (iv) division of epithelial cells into daughter cells was circumferential and not radial; (v) bromodeoxyuridine-labelled basal cell nuclei were mostly not found in the vicinity of labelled principal cell nuclei and vice versa; and (vi) the percentage of labelled basal cell nuclei was higher than that of the other cells, which is indicative of their arrival from a more highly labelled pool. In addition, no age-dependent correlation was noted between any of the cytokines tested and appearance of basal cells in the epithelium, and basal cells expressed intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1), which provides further evidence of a relationship between basal cells and immunocytes. If basal cells have an immunological function, failure of their recruitment into the epididymal epithelium at about the time of puberty may have repercussions for immunological protection of spermatozoa and, ultimately, for fertility in the adult.


Exacerbation of herpes simplex encephalitis after successful treatment with acyclovir.
Herpes simplex encephalitis (HSE) in children sometimes exacerbates after successful treatment; yet the frequency, etiology, and clinical features of exacerbation remain unclear. We report data for 27 children with HSE confirmed by polymerase chain reaction (PCR) analysis; all were successfully treated with acyclovir, but 7 (26%) had a relapse of encephalitic illness. In 2 of those 7, serial examination with a PCR assay showed that herpes simplex virus (HSV) DNA reappeared temporarily in the cerebrospinal fluid (CSF). For 5 of the 7 patients, a second course of acyclovir therapy was effective. Coxsackievirus A9 was isolated from CSF of 1 case patient during subsequent exacerbation. The total dose during initial acyclovir therapy was significantly lower in the relapse group than in the control group (P=.027). In conclusion, exacerbation of HSE in children may be more common than previously recognized. It is suggested that the replication of HSV or another viral pathogen caused a second encephalitic illness (HSE) in some cases.


Dosage and timing of anti-TNF-alpha antibody treatment determine its effect of resistance to sepsis after injury.
Antibody against tumor necrosis factor-alpha (TNF-alpha) has improved survival in certain models of sepsis, but it remains unproven in clinical studies. In most of the successful animal studies, efficacy has been shown in previously healthy animals subjected to a septic challenge. Patients at risk for sepsis, however, may be ill for some time before the sepsis supervenes. This situation has been described as a "two-hit" model of critical illness. We have developed an animal burn-sepsis model which conforms to this "two-hit" concept. We have quantified macrophage TNF-alpha production at different times after the burn (first "hit") and determined the effect of neutralizing antibody against TNF-alpha during this period on survival after subsequent sepsis (second "hit"). The objective of this study was to determine the role of TNF-alpha and the effect of neutralizing antibody against TNF-alpha in a burn-sepsis model. Animals were subjected to a full thickness burn or sham burn. In vitro TNF-alpha production from cultured lipopolysaccharide-stimulated splenic adherent cells was determined at various time points thereafter by enzyme-linked immunosorbent assay. Separate animals were treated with neutralizing antibody against TNF-alpha at different time points after the thermal injury, and survival was determined after septic challenge (cecal ligation and puncture) on Day 10 after the burn. TNF-alpha production from adherent splenocytes was not elevated in the early days after thermal injury, but was significantly enhanced from Day 6 onward compared with sham-burned animals. Nine percent of the burned mice survived septic challenge compared with 69% of the sham-burned control mice (P < 0.001). Therapy with anti-TNF antibody at 1 x 10(4) neutralizing units (n.u.) kg-1 markedly improved outcome if given when TNF-alpha production was elevated at Day 7 after the burn (survival, 36%; P = 0.01) but did not improve survival when administered at Days 0 or 4 or at the time of the septic challenge (Day 10). High doses of antibody (3.2 x 10(5) n.u.kg-1) were not beneficial and may have been detrimental. These results show that neutralizing antibody against TNF-alpha may reduce the susceptibility to infection seen after thermal injury, but the timing of administration of the antibody and the dose of antibody used are critical to the outcome. This should be considered when neutralizing antibody against TNF is used in the clinical setting.


Capillary-Induced Hair Twist.
We investigate the self-assembly of hair-like fibers into twisted helices as they are pulled through the liquid interface at a controlled rate. Capillary-induced spontaneous fiber twisting phenomena are observed from the nano- to the millimeter scale. Here, we control the drain rate of the liquid and observe two regimes of self-assembly of long hairs. At low drain rates, the hairs coalesce radially to form a dense aggregate. At higher drain rates, spontaneous hair twisting occurs. We find that the drain rate corresponding to the twisting threshold scales with the characteristic velocity of fiber coalescence set by a balance between liquid viscosity μ and surface energy σ and reads ∼(σ/μ)·(S/l)2 where S and l are the spacing between hairs and their length, respectively. At drain rates higher than this threshold, liquid is entrained between the hairs as they emerge from the liquid surface, forming a circular liquid column. Twisting is induced by the fast radial shrinking of this liquid column, combined with the nonlinear resistance to the hairs' radial versus tangential coalescence. Understanding the kinetics is crucial to control this complex self-assembly and to engineer fiber drying processes at various length scales.


Acute Gout in Hospitalized patients in Sarawak General Hospital.
we performed a prospective study of all hospitalized patients with a diagnosis of Gout in Sarawak General hospital from 1st July 2011 to 1st July 2012. There were a total of 126 patients in our study of which 112 (88.9%) were males. The majority of our patients were from the indigenous populations (71.7%). They have a mean age of 60.0 ± 14.2 years. Most of our patients were overweight (68%) with comorbities of hypertension (78.6%), Chronic Kidney Failure (48.4%), Type II diabetes Mellitus (30.2%), dyslipidemia (27.8%) and Ischaemic heart disease (11.9%). Polyarticular gouty arthritis was the main presenting pattern during hospitalization (88.1%). The mean length of stay for our patients was 9.8 ± 6.0 days which was significantly longer than the mean length of stay for other patients without gout (p<0.05). Only 17 patients had gout on admission and the majority developed gout during hospitalizations. Our patients were admitted respectively for medical problems (45.4%), surgical problems (28.6%) and orthopaedic problems (9.2%). Colchicine (73.8%) and steroid (40.5%) were the main stays of treatment for our patients. Our hospitalized gout patients were complicated patients with multiple comorbidities.


Assessment of Implicit Gender Bias During Evaluation of Procedural Competency Among Emergency Medicine Residents.
Gender disparities exist throughout medicine. Recent studies have highlighted an attainment gap between male and female residents in performance evaluations on Accreditation Council for Graduate Medical Education (ACGME) milestones. Because of difficulties in blinding evaluators to gender, it remains unclear whether these observed disparities are because of implicit bias or other causes. To estimate the magnitude of implicit gender bias in assessments of procedural competency in emergency medicine residents and whether the gender of the evaluator is associated with identified implicit gender bias. A cross-sectional study was performed from 2018 to 2020 in which emergency medicine residency faculty assessed procedural competency by evaluating videos of residents performing 3 procedures in a simulated environment. They were blinded to the intent of the study. Proceduralists were filmed performing each procedure from 2 different viewpoints simultaneously by 2 different cameras. One was a gender-blinded (ie, hands-only) view, and the other a wide-angled gender-evident (ie, whole-body) view. The faculty evaluators viewed videos in a random order and assessed procedural competency on a global rating scale with extensive validity evidence for the evaluation of video-recorded procedural performance. The primary outcome was to determine if there was a difference in the evaluation of procedural competency based on gender. The secondary outcome was to determine if there was a difference in the evaluations based on the gender of the evaluator. Fifty-one faculty evaluators enrolled from 19 states, with 22 male participants (43.1%), 29 female participants (56.9%), and a mean (SD) age of 37 (6.4) years. Each evaluator assessed all 60 procedures: 30 gender-blinded (hands-only view) videos and 30 identical gender-evident (wide angle) videos. There were no statistically significant differences in the study evaluators' scores of the proceduralists based on their gender, and the gender of the evaluator was not associated with the difference in mean scores. In this study, we did not identify a difference in the evaluation of procedural competency based upon the gender of the resident proceduralist or the gender of the faculty evaluator.


Impact of Remote and Virtual Care Models on the Sustainability of Small Health Care Businesses: Perceptual Analysis of Small Clinics, Physician Offices, and Pharmacies in Colorado.
Lockdowns and shelter-in-place orders during COVID-19 have accelerated the adoption of remote and virtual care (RVC) models, potentially including telehealth, telemedicine, and internet-based electronic physician visits (e-visits) for remote consultation, diagnosis, and care, deterring small health care businesses including clinics, physician offices, and pharmacies from aligning resources and operations to new RVC realities. Current perceptions of small health care businesses toward remote care, particularly perceptions of whether RVC adoption will synergistically improve business sustainability, would highlight the pros and cons of rapidly adopting RVC technology among policy makers. This study aimed to assess the perceptions of small health care businesses regarding the impact of RVC on their business sustainability during COVID-19, gauge their perceptions of their current levels of adoption of and satisfaction with RVC models and analyze how well that aligns with their perceptions of the current business scenario (SCBS), and determine whether these perceptions influence their view of their midterm sustainability (SUST). We randomly sampled small clinics, physician offices, and pharmacies across Colorado and sought assistance from a consulting firm to collect survey data in July 2020. Focal estimated study effects were compared across the three groups of small businesses to draw several insights. In total, 270 respondents, including 82 clinics, 99 small physician offices, and 89 pharmacies, across Colorado were included. SRVC and SCBS had direct, significant, and positive effects on SUST. However, we investigated the effect of the interaction between SRVC and SCBS to determine whether RVC adoption aligns with their perceptions of the current business scenario and whether this interaction impacts their perception of business sustainability. Effects differed among the three groups. The interaction term SRVC×SCBS was significant and positive for clinics (P=.02), significant and negative for physician offices (P=.05), and not significant for pharmacies (P=.76). These variations indicate that while clinics positively perceived RVC alignment with the current business scenario, the opposite held true for small physician offices. As COVID-19 continues to spread worldwide and RVC adoption progresses rapidly, it is critical to understand the impact of RVC on small health care businesses and their perceptions of long-term survival. Small physician practices cannot harness RVC developments and, in contrast with clinics, consider it incompatible with business survival during and after COVID-19. If small health care firms cannot compete with RVC (or synergistically integrate RVC platforms into their current business practices) and eventually become nonoperational, the resulting damage to traditional health care services may be severe, particularly for critical care delivery and other important services that RVC cannot effectively replace. Our results have implications for public policy decisions such as incentive-aligned models, policy-initiated incentives, and payer-based strategies for improved alignment between RVC and existing models.


Triggered drug release from dynamic microspheres via a protein conformational change.
In this study we formed and characterized dynamic hydrogel microspheres in which a protein conformational change was used to control microsphere volume changes and the release of an encapsulated drug. In particular, a specific biochemical ligand, trifluoperazine, induced calmodulin's nanometer scale conformation change, which translated to a 48.7% microsphere volume decrease. This specific, ligand-induced volume change triggered the release of a model drug, vascular endothelial growth factor (VEGF), at pre-determined times. After release from the microspheres, 85.6 +/- 10.5% of VEGF was in its native conformation. Taken together, these results suggest that protein conformational change could serve as a useful mechanism to control drug release from dynamic hydrogels.


Auditory cortical activity after intracortical microstimulation and its role for sensory processing and learning.
Several studies have shown that animals can learn to make specific use of intracortical microstimulation (ICMS) of sensory cortex within behavioral tasks. Here, we investigate how the focal, artificial activation by ICMS leads to a meaningful, behaviorally interpretable signal. In natural learning, this involves large-scale activity patterns in widespread brain-networks. We therefore trained gerbils to discriminate closely neighboring ICMS sites within primary auditory cortex producing evoked responses largely overlapping in space. In parallel, during training, we recorded electrocorticograms (ECoGs) at high spatial resolution. Applying a multivariate classification procedure, we identified late spatial patterns that emerged with discrimination learning from the ongoing poststimulus ECoG. These patterns contained information about the preceding conditioned stimulus, and were associated with a subsequent correct behavioral response by the animal. Thereby, relevant pattern information was mainly carried by neuron populations outside the range of the lateral spatial spread of ICMS-evoked cortical activation (approximately 1.2 mm). This demonstrates that the stimulated cortical area not only encoded information about the stimulation sites by its focal, stimulus-driven activation, but also provided meaningful signals in its ongoing activity related to the interpretation of ICMS learned by the animal. This involved the stimulated area as a whole, and apparently required large-scale integration in the brain. However, ICMS locally interfered with the ongoing cortical dynamics by suppressing pattern formation near the stimulation sites. The interaction between ICMS and ongoing cortical activity has several implications for the design of ICMS protocols and cortical neuroprostheses, since the meaningful interpretation of ICMS depends on this interaction.


Hyperbolic Modeling of Subthalamic Nucleus Cells to Investigate the Effect of Dopamine Depletion.
To investigate how different types of neurons can produce well-known spiking patterns, a new computationally efficient model is proposed in this paper. This model can help realize the neuronal interconnection issues. The model can demonstrate various neuronal behaviors observed in vivo through simple parameter modification. The behaviors include tonic and phasic spiking, tonic and phasic bursting, class 1 and class 2 excitability, rebound spike, rebound burst, subthreshold oscillation, and accommodated spiking along with inhibition neuron responses. Here, we investigate the neuronal spiking patterns in Parkinson's disease through our proposed model. Abnormal pattern of subthalamic nucleus in Parkinson's disease can be studied through variations in the shape and frequency of firing patterns. Our proposed model introduces mathematical equations, where these patterns can be derived and clearly differentiated from one another. The irregular and arrhythmic behaviors of subthalamic nucleus firing pattern under normal conditions can easily be transformed to those caused by Parkinson's disease through simple parameter modifications in the proposed model. This model can explicitly show the change of neuronal activity patterns in Parkinson's disease, which may eventually lead to effective treatment with deep brain stimulation devices.


Neuronal activity of the human subthalamic nucleus in the parkinsonian and nonparkinsonian state.
We recorded resting-state neuronal activity from the human subthalamic nucleus (STN) during functional stereotactic surgeries. By inserting up to five parallel microelectrodes for single- or multiunit recordings and applying statistical spike-sorting methods, we were able to isolate a total of 351 single units in 65 patients with Parkinson's disease (PD) and 33 single units in 9 patients suffering from essential tremor (ET). Among these were 93 pairs of simultaneously recorded neurons in PD and 17 in ET, which were detected either by the same (n = 30) or neighboring microelectrodes (n = 80). Essential tremor is a movement disorder without any known basal ganglia pathology and with normal dopaminergic brain function. By comparing the neuronal activity of the STN in patients suffering from PD and ET we intended to characterize, for the first time, changes of basal ganglia activity in the human disease state that had previously been described in animal models of Parkinson's disease. We found a significant increase in the mean firing rate of STN neurons in PD and a relatively larger fraction of neurons exhibiting burstlike activity compared with ET. The overall proportion of neurons exhibiting intrinsic oscillations or interneuronal synchronization as defined by significant spectral peaks in the auto- or cross-correlations functions did not differ between PD and ET when considering the entire frequency range of 1-100 Hz. The distribution of significant oscillations across the theta (1-8 Hz), alpha (8-12 Hz), beta (12-35 Hz), and gamma band (>35 Hz), however, was uneven in ET and PD, as indicated by a trend in Fisher's exact test (P = 0.05). Oscillations and pairwise synchronizations within the 12- to 35-Hz band were a unique feature of PD. Our results confirm the predictions of the rate model of Parkinson's disease. In addition, they emphasize abnormalities in the patterning and dynamics of neuronal discharges in the parkinsonian STN, which support current concepts of abnormal motor loop oscillations in Parkinson's disease.


Determinants of pancreatic microcirculation in acute pancreatitis in rats.
The aim of the study was to evaluate the effects of arterial hypotension, high-volume crystalloid resuscitation, and isovolemic hemodilution on pancreatic microvascular perfusion during acute pancreatitis. Using intravital microscopy, pancreatic functional capillary density was analyzed in rats 1 and 2 hr after onset of acute pancreatitis. Pancreatic microvascular perfusion in acute pancreatitis was characterized by a (-62%) significant reduction of functional capillary density predominantly in perinecrotic but also in nonnecrotic tissue (-43%). Pancreatic microvascular perfusion failure was aggravated by arterial hypotension but attenuated by treatment with high-volume crystalloid resuscitation. Isovolemic hemodilution was found superior to high-volume crystalloid resuscitation in maintaining pancreatic functional capillary density and therefore has the best potential in preserving tissue integrity and thereby limiting progression of disease. This study underlines the importance of early fluid resuscitation/hemodilution in patients presenting with acute pancreatitis.


LPA-primed astrocytes induce axonal outgrowth of cortical progenitors by activating PKA signaling pathways and modulating extracellular matrix proteins.
Lysophosphatidic acid (LPA) is one of the main membrane-derived lysophospholipids, inducing diverse cellular responses like cell proliferation, cell death inhibition, and cytoskeletal rearrangement, and thus is important in many biological processes. In the central nervous system (CNS), post-mitotic neurons release LPA extracellularly whereas astrocytes do not. Astrocytes play a key role in brain development and pathology, producing various cytokines, chemokines, growth factors, and extracellular matrix (ECM) components that act as molecular coordinators of neuron-glia communication. However, many molecular mechanisms underlying these events remain unclear-in particular, how the multifaceted interplay between the signaling pathways regulated by lysophospholipids is integrated in the complex nature of the CNS. Previously we showed that LPA-primed astrocytes induce neuronal commitment by activating LPA1-LPA2 receptors. Further, we revealed that these events were mediated by modulation and organization of laminin levels by astrocytes, through the induction of the epidermal growth factor receptor (EGFR) signaling pathway and the activation of the mitogen-activated protein (MAP) kinase (MAPK) cascade in response to LPA (Spohr et al., 2008, 2011). In the present work, we aimed to answer whether LPA affects astrocytic production and rearrangement of fibronectin, and to investigate the mechanisms involved in neuronal differentiation and maturation of cortical neurons induced by LPA-primed astrocytes. We show that PKA activation is required for LPA-primed astrocytes to induce neurite outgrowth and neuronal maturation and to rearrange and enhance the production of fibronectin and laminin. We propose a potential mechanism by which neurons and astrocytes communicate, as well as how such interactions drive cellular events such as neurite outgrowth, cell fate commitment, and maturation.


Visceral leishmaniasis affects liver and spleen concentrations of amphotericin B following administration to mice.
To assess the impact of visceral leishmaniasis (VL) on the concentration of amphotericin B (AmB) recovered in the liver and spleen following either intravenous (AmBisome) or oral (iCo-009) AmB administration to mice. Livers and spleens previously obtained from VL-infected BALB/c mice (following intravenous AmBisome or oral AmB treatments) were analysed for AmB concentrations. Then, non-infected BALB/c mice were divided into three treatment groups: a single dose of intravenous AmBisome (2 mg/kg, n = 5); and oral AmB every 12 h for 5 days (10 mg/kg, n = 6 and 20 mg/kg, n = 6). The animals were sacrificed 7 days after the initiation of the treatment and the livers and spleens were harvested for drug analysis by HPLC. The single intravenous injection of AmBisome resulted in a 77-fold lower concentration of AmB in infected compared with non-infected liver tissue, while the difference in AmB concentration in the spleen was only 5-fold. The multiple dose oral administration of AmB resulted in a 3-fold lower concentration of AmB in infected compared with non-infected livers for both oral doses, while the differences in AmB concentrations in the spleen were not statistically different for the oral treatment groups. VL significantly lowered the concentration of AmB in the liver and the spleen when compared with uninfected animals. This effect seems to correlate with the degree of infection of the tissue. In the case of the intravenous liposomal formulation (AmBisome), the differences between the infected and non-infected tissues are of a higher magnitude than in the case of orally administered AmB (iCo-009).


Synthesis, Characterization, and Density Functional Theory Analysis of Uranium and Thorium Complexes Containing Nitrogen-Rich 5-Methyltetrazolate Ligands.
Two nitrogen-rich, isostructural complexes of uranium and thorium, (C5Me5)2U[η(2)-(N,N')-tetrazolate]2 (7) and (C5Me5)2Th[η(2)-(N,N')-tetrazolate]2 (8), containing 5-methyltetrazolate, have been synthesized and structurally characterized by single-crystal X-ray diffraction, electrochemical methods, UV-visible-near-IR spectroscopy, and variable-temperature (1)H NMR spectroscopy. Density functional theory (DFT) calculations yield favorable free energies of formation (approximately -375 kJ/mol) and optimized structures in good agreement with the experimental crystal structures. Additionally, calculated NMR chemical shifts of 7 and 8 are in good agreement with the variable-temperature (1)H NMR experiments. Time-dependent DFT calculations of both complexes yield UV-visible spectroscopic features that are consistent with experiment and provide assignments of the corresponding electronic transitions. The electronic transitions in the UV-visible spectroscopic region are attributed to C5Me5 ligand-to-metal charge transfer. The low-lying molecular orbitals of the tetrazolate ligands (∼2 eV below the HOMO) do not contribute appreciably to experimentally observed electronic transitions. The combined experimental and theoretical analysis of these new nitrogen-rich uranium and thorium complexes indicates the tetrazolate ligand behaves primarily as a σ-donor.


Neck injury is critical to elastase-induced aneurysm model.
We modified the elastase-induced aneurysm model by use of a simple surgical technique in rabbits. A temporary arcuated aneurysm clip was placed at the origin of the right common carotid artery (RCCA), ascertaining the inner edge of the clip blade on the juncture of the RCCA and right subclavian artery (RSCA), and the elastase-induced aneurysm procedure was undertaken. We found elastase and location of the temporary arcuated aneurysm clip are critical to the success of this aneurysm model.


Inhibiting PHD2 in bone marrow mesenchymal stem cells via lentiviral vector-mediated RNA interference facilitates the repair of periodontal tissue defects in SD rats.
Hypoxia-inducible factors (HIFs) play an important role in angiogenesis, and they can activate the expression of several downstream angiogenic factors. HIF-1 is a major transcriptor of HIFs, composed of α and β subunits. Prolyl hydroxylase domain-containing protein 2 (PHD2) is the main catabolic enzyme for HIF-1α, and it can accelerate its degradation under normoxic conditions. PHD2 expression in bone marrow mesenchymal stem cells (BMMSCs) of SD rats was down-regulated under normoxic conditions in this study by utilizing lentiviral vector-mediated RNA interference to promote HIF-1α accumulation, thus enhancing the expression of angiogenic factors. A tissue-engineered compound was constructed using the composite collagen membrane of BMMSCs after PHD2 gene silencing to repair periodontal fenestration defects in SD rats. The results of this study indicated that, after PHD2 gene silencing, the osteogenic differentiation of BMMSCs was enhanced in vitro, the resistance of cells to oxidative stress was also validated in vitro, thereby illustrating the promotion of the repair of artificially constructed periodontal tissue defects in rats. The results of this study provide a reference and guidance for future applications of RNA interference in periodontal tissue engineering and serve as a basis for improving the survival of seed cells in recipient tissues.


Streptococcal toxic shock syndrome manifesting as peritonitis in a child.
Streptococcal toxic shock syndrome (TSS) with the initial manifestation of peritonitis is rare. We report the case of a 5-year-old girl who presented with peritonitis and shock. Emergency laparotomy was performed but no perforated visceral organ was found. Acute respiratory distress syndrome, impaired renal function, and coagulopathy developed later. Group A beta-hemolytic Streptococcus (GAS) was isolated from the pleural effusion and the diagnosis of streptococcal TSS was made. This association highlights the need for thorough examination and close observation in the management of childhood peritonitis.


[Gene cloning, codon optimization and functional expression of Yarrawia lipolytica lipase Lip1].
To implement inducible and constitutive over-expression of Yarrowia lipolytica lipase gene lipl in Pichia pastoris using codon optimization. We cloned Y. lipolytica lipase gene lip1 according to codon bias of P. pastoris, and optimized lipl using overlap extension PCR synthesis. Then, we cloned the original and optimized genes into the induced vector pPIC9K and newly built constitutive carrier pGAP9K, and electrotransformated the resultant expression plasmids into P. pastoris GS115. Through G418 resistance screening, high copy transformatants were selected and fermented in shake flasks. P-nitrophenyl palmitate (pNPP) was used as substrates for assay the activities of the expressed lipase, and the characteristics of the lipase were further examined. We successfully cloned lipase gene lip1 from Y. lipolytica, nucleotide sequence revealed that the open reading frame (ORF) had 1461 nucleotides, encoding 486 amino acids, without any intron or any signal peptide. SDS-PAGE analysis and fermentation result showed that the optimized gene had a higher expression level than the original one, and the constitutive expression was superior to the inducible expression. Preliminary analysis showed that the optimal substrate of Lipl was p-nitrophenyl butyrate (C4), the optimum temperature and pH was 45 degrees C and 8.5, respectively. Y. lipolytica lipase gene lip1 can be over-expressed through both inducible and constitutive expressions using codon optimization, which lays a solid foundation to further study Y. lipolytica lipase family, and also provides an important prerequisite for scale production and industrial application of the lipase.


Creep rupture of viscoelastic fiber bundles.
We study the creep rupture of bundles of viscoelastic fibers occurring under uniaxial constant tensile loading. A fiber bundle model is introduced that combines the viscoelastic constitutive behavior and the strain controlled breaking of fibers. Analytical and numerical calculations showed that above a critical external load the deformation of the system monotonically increases in time resulting in global failure at a finite time t(f), while below the critical load the deformation tends to a constant value giving rise to an infinite lifetime. Our studies revealed that the nature of the transition between the two regimes, i.e., the behavior of t(f) at the critical load sigma(c), strongly depends on the range of load sharing: for global load sharing t(f) has a power law divergence at sigma(c) with a universal exponent of 0.5, however, for local load sharing the transition becomes abrupt: at the critical load t(f) jumps to a finite value, analogous to second- and first-order phase transitions, respectively. The acoustic response of the bundle during creep is also studied.