The NCLX-type Na+/Ca2+ Exchanger NCX-9 Is Required for Patterning of Neural Circuits in Caenorhabditis elegans.
NCLX is a Na+/Ca2+ exchanger that uses energy stored in the transmembrane sodium gradient to facilitate the exchange of sodium ions for ionic calcium. Mammals have a single NCLX, which has been shown to function primarily at the mitochondrion and is an important regulator of neuronal physiology by contributing to neurotransmission and synaptic plasticity. The role of NCLX in developmental cell patterning (e.g. in neural circuits) is largely unknown. Here we describe a novel role for the Caenorhabditis elegans NCLX-type protein, NCX-9, in neural circuit formation. NCX-9 functions in hypodermal seam cells that secrete the axon guidance cue UNC-129/BMP, and our data revealed that ncx-9-/- mutant animals exhibit development defects in stereotyped left/right axon guidance choices within the GABAergic motor neuron circuit. Our data also implicate NCX-9 in a LON-2/heparan sulfate and UNC-6/netrin-mediated, RAC-dependent signaling pathway to guide left/right patterning within this circuit. Finally, we also provide in vitro physiology data supporting the role for NCX-9 in handling calcium exchange at the mitochondrion. Taken together, our work reveals the specificity by which the handling by NCLX of calcium exchange can map to neural circuit patterning and axon guidance decisions during development.


Mechanisms of response to silver nanoparticles on Enchytraeus albidus (Oligochaeta): survival, reproduction and gene expression profile.
Silver has antimicrobial properties and silver nanoparticles (Ag-NPs) have been some of the most widely used NPs. Information regarding their effects is still insufficient, in particular for soil dwelling organisms. The standard soil Oligochaete Enchytraeus albidus was used to study the effects of Ag in soils, using differential gene expression (microarray) and population (survival, reproduction) response to Ag-NPs (PVP coated) and AgNO₃. Results showed higher toxicity of AgNO₃ (EC₅₀<50 mg/kg) compared to toxicity of Ag-NPs (EC₅₀=225 mg/kg). Based on the biological and material identity, the difference in toxicity between Ag-NPs and AgNO₃ could possibly be explained by a release of Ag(+) ions from the particles or by a slower uptake of Ag-NPs. The indications were that the responses to Ag-NPs reflect an effect of Ag ions and Ag-NPs given the extent of similar/dissimilar genes activated. The particles characterization supports this deduction as there were limited free ions measured in soil extracts, maybe related to little oxidation and/or complexation in the soil matrix. The possibility that gene differences were due to different levels of biological impact (i.e. physiological responses) should not be excluded. Testing of Ag-NPs seem to require longer exposure period to be comparable in terms of effect/risk assessment with other chemicals.


A reconstituted in vitro clot model for evaluating laser thrombolysis.
Laser thrombolysis is the selective removal of thrombus from occluded blood vessels using laser energy. A reconstituted clot model with reproducible optical absorption properties was developed to evaluate the effect of various laser parameters on thrombus removal rate. Reconstituted clots were made with known fibrinogen concentrations and hematocrits. Ex vivo clots were collected from ten swine. Four red gelatin phantoms were prepared. Mass removal rates and ablation efficiencies were determined using a 577 nm, 1 microsec pulsed dye laser. The ablation efficiencies of the three clot models were compared at an energy of 25 mJ and a repetition rate of 4 Hz. In addition, the reconstituted clot model was ablated as pulse energy and repetition rate were varied with average power held constant at 100 mW. The mean ablation efficiency for ex vivo clots ranged from 0.4 +/- 0.1 to 3.4 +/- 0.7 microg/mJ/pulse, with significant differences between groups (ANOVA p < 0.05). Reconstituted clots of varied fibrinogen content had ablation efficiencies of 1.5 +/- 0.2 to 1.6 +/- 0.3 microg/mJ/pulse at this energy and repetition rate. Gelatin ablation efficiency was inversely proportional to protein content and ranged from 0.5 +/- 0.3 to 2.0 +/- 0.7 microg/mJ/pulse. Reconstituted clot mass removal rates (in microg/s) were clinically similar for settings ranging from 13 mJ at 8 Hz to 33 mJ at 3 Hz. The reconstituted model clot is a reproducible and biologically relevant thrombolysis target. Ex vivo clot lacks reproducibility between individuals and gelatin phantoms lack clinical relevance. At a constant average power, varying laser parameters did not affect mass removal rates to a clinically significant degree.


Functional exploration of the IFT-A complex in intraflagellar transport and ciliogenesis.
Intraflagellar transport (IFT) particles or trains are composed of IFT-A and IFT-B complexes. To assess the working mechanism of the IFT-A complex in IFT and ciliogenesis, we have analyzed ift43 mutants of Chlamydomnonas in conjunction with mutants of the other IFT-A subunits. An ift43 null mutant or a mutant with a partial deletion of the IFT43 conserved domain has no or short flagella. The mutants accumulate not only IFT-B but also IFT-Ain the short flagella, which is in contrast to an ift140 null mutant. The IFT43 conserved domain is necessary and sufficient for the function of IFT43. IFT43 directly interacts with IFT121 and loss of IFT43 results in instability of IFT-A. A construct with a partial deletion of the IFT43 conserved domain is sufficient to rescue the instability phenotype of IFT-A, but results in diminishing of IFT-A at the peri-basal body region. We have further provided evidence for the direct interactions within the IFT-A complex and shown that the integrity of IFT-A is important for its stability and cellular localization. Finally, we show that both IFT43 and IFT140 are involved in mobilizing ciliary precursors from the cytoplasmic pool during flagellar regeneration, suggesting a novel role of IFT-A in transporting ciliary components in the cytoplasm to the peri-basal body region.


Open channel block of the fast transient outward K+ current by primaquine and chloroquine in rat left ventricular cardiomyocytes.
Anti-malarial drugs may have severe adverse cardiac effects as a result of their ion channel blocking properties. Here we investigate the effect of the aminoquinolines primaquine and chloroquine on the fast transient outward K(+) current (I(to)) of single epicardial myocytes isolated from the left ventricular free wall of female Wistar rats. The ruptured-patch whole-cell configuration of the patch-clamp technique was used to investigate I(to). At +60 mV, primaquine blocked I(to) amplitude (defined as the current inactivating during a test pulse of 600 ms duration) with an IC(50) of 118±8 μM. I(to) charge was blocked with an IC(50) of 33±2 μM (n=42), indicating open channel block. Chloroquine blocked I(to) amplitude with an IC(50) of 4.6±0.9 mM, while the IC(50) for I(to) charge was 439±63 μM (n=23). The kinetic analysis of the onset of block revealed K(d) values of 52±8 μM (n=18) and 520±60μM (n=11) for primaquine and chloroquine, respectively. Both drugs significantly accelerated the apparent inactivation time constant of I(to). Steady-state inactivation of I(to) was not altered by 30 μM primaquine. In contrast, I(to) recovery from inactivation was prolonged with the appearance of an additional long time constant without a change of the short time constant. Exposure to 1mM chloroquine resulted in a right shift of steady-state inactivation, whereas recovery from inactivation was only mildly affected. Both substances exhibited considerable use dependence. In X. laevis oocytes heterologously expressing hKv4.2+hKChIP2b channels the block by the aminoquinolines was voltage dependent. We conclude that primaquine and chloroquine are open-channel blockers of I(to).


[Research progress of dental pulp stem cells for peripheral nerve injury repair].
Peripheral nerve injuries are mainly related to severe trauma, fracture and tumor surgery, leading to reduced quality of life and impaired physical and mental health. The repair of peripheral nerve still faces great challenges in clinic, and the research on the regeneration and repair of peripheral nerve has become a hot issue in related disciplines. Cell therapy plays an irreplaceable role in tissue regeneration and repair. Schwann cells are ideal cells for peripheral nerve repair, but their limited sources inhibit the clinical application. Dental pulp stem cells are derived from neural crest, which provides a new cell source for nerve regeneration. The purpose of this article is to review the research progress of dental pulp stem cells for peripheral nerve repair.


[Diagnosis related groups (DRG) in child and adolescent psychiatry: results of a prospective pilot study].
Germany faces one of the biggest 'revolutions' in the health care system. As decided by the government in cooperation with medical boards and insurances, beginning with January 1st, 2003 all inpatient treatments will be paid on the basis of adapted Australian-Refined Diagnosis Related Groups. To date, hospitals are requested to obtain prospective databases in order to cluster homogeneous diagnostic groups and calculate realistic treatment-costs. Both psychiatry and child and adolescent psychiatry are so far excluded from the introduction of DRGs. However, most experts predict extensive shifts of patients into psychiatry (i.e. with comorbid internal diseases) under the pressure of short treatments in all non-psychiatric disciplines. Therefore, changes in the payment of psychiatric treatments are inevitably. As part of the DRG pilot-study, we created a catalogue adapted to child and adolescent psychiatry, which was used for 102 consecutively treated inpatients of a child and adolescent hospital. A total of 17.019 prospectively assessed procedures were obtained. Under clinical aspects, 11 categories of 'typical' disorders were analysed. Worst predictability of treatment costs was found for obsessive-compulsive disorders, personality disorders, and eating disorders. Comorbidity and complexity of the disorder was not related to the length of hospital treatment. Implications on future payment systems in child and adolescent psychiatry are discussed.


Optimising waiting: a view from the English National Health Service.
Recent authors have proposed that waiting times for elective treatment should be reduced to the point where the costs of doing so exceed the benefits. This paper considers how this criterion could be put into effect. Taking benefits first it argues that these could be estimated in three different ways - social cost benefit, clinical and user valuation - that would not necessarily produce consistent results and hence a choice has to be made between them. It then considers the costs of reducing waits and argues, citing relevant evidence, that these may range widely according to whether or not reductions can be achieved through simple management measures or whether more long-term capacity is required. It concludes therefore that the apparently simple criterion proposed for defining the point where waiting times are optimal is hard to establish. Choice of criterion must be made in the light of the overall values that a given health care system is intended to promote.


Computed tomography assessment of early response to neoadjuvant therapy in colon cancer.
Using multidetector computed tomography, we aimed to assess the early response of neoadjuvant drug therapy for locally advanced colon cancer. Computed tomography with IV contrast was acquired from 67 patients before and after up to three cycles of preoperative treatment. All patients had histologically confirmed colon cancer, a T4 or T3 tumour with extramural invasion ≥ 5 mm and no distant metastases or peritoneal nodules. The patients were treated with oxaliplatin and capecitabine. In addition, those with no mutations in the KRAS, BRAF and PIK3CA genes were also treated with panitumumab. Before and after treatment, we measured the tumour diameter in two different planes, the extension of the extramural tumour invasion, and the number and size of enlarged lymph nodes. The mean tumour length was 7.8 cm (95% confidence interval (CI): 5.3-10.4) at baseline and 4.34 cm (95% CI: 4.0-4.9) after treatment. The mean extramural tumour invasion was 10.6 mm (95% CI: 9.5-11.8) at baseline and 5.7 mm (95% CI: 4.7-6.7) after treatment. The mean number of enlarged lymph nodes was 4.1 (95% CI: 3.4-4.9) at baseline and 2.1 (95% CI: 1.4-2.7) after treatment. According to RECIST 1.1, 45% (95% CI: 34-57) of the patients had a response and 55% (95% CI: 43-67) had stable disease. None of the patients showed progressive disease. Using CT, we demonstrated a significant reduction in tumour size, extramural tumour invasion, number and size of enlarged lymph nodes following neoadjuvant treatment. not relevant. Registered with ClinicalTrials.gov (NCT 01108107).


Benefit-incidence analysis of government spending on Ministry of Health outpatient services in Jordan.
A benefit-incidence analysis was conducted for the year 2000 using various data sources including the Jordan healthcare utilization and expenditure survey 2000. The results illustrate that the poorest segment of the Jordanian population were the most likely to report sickness and seek treatment and were the main users of the Ministry of Health outpatient services. The poorest uninsured individuals were the main source of revenues generated through user fees. The targeting efficiency (i.e. total percentage of benefits received) for the poorest quintile was 33.8% compared with 4.0% in the richest quintile. The analysis demonstrates that the Jordanian government in-kind subsidy is reaching the poor.


Steroids modulate transepithelial resistance and Na(+) absorption across cultured porcine vas deferens epithelia.
Epithelial cells were isolated from adult porcine vas deferens and grown in the absence or presence of steroid hormones. Transepithelial resistance (R(te)), basal short circuit current (I(sc)), and the effects of selected ion transport modulators on these parameters were evaluated in modified Ussing chambers at three time points (5-8, 11-14, and 18-22 days postseeding). At the earliest time point, no significant differences were observed. At the middle time point, when compared with R(te) in untreated control monolayers, R(te) in monolayers exposed to 17beta-estradiol, aldosterone, cortisol, cortisone, prednisolone, prednisone, and dexamethasone was significantly lower; in contrast, R(te) in monolayers exposed to testosterone, dihydrotestosterone, or progesterone did not differ from that in control monolayers. Treatments with cortisol, prednisolone, and dexamethasone were associated with an elevated basal I(sc) that was amiloride sensitive, indicating ongoing Na(+) absorption by these monolayers. R(te) was increased by amiloride treatment in glucocorticoid-treated monolayers but remained significantly less than that of control monolayers. At the third time point, the postamiloride R(te) of glucocorticoid-treated monolayers was not different from that of control monolayers. Responses to ATP, forskolin, bumetanide, and DASU-02 were not affected by steroid treatment at any time point. Taken together, these results suggest that estrogens and corticosteroids can modulate epithelial function in the distal excurrent duct of the adult male reproductive system. At physiological or pharmacological concentrations, these hormones would be expected to modify the luminal environment (both the ionic composition and pH) to which sperm are exposed and thus affect male fertility.


Case and reanalysis.
In this paper we discuss an asymmetry in the Case system of German and its implications for human sentence processing: the asymmetry between nominative/accusative and dative case. Starting from the assumption that dative case has a distinct grammatical representation--dative DPs are embedded into an extra structural layer KP--the results of two experiments will be presented, which show that dative assignment during reanalysis is accompanied by additional processing operations that are not needed when accusative or nominative are assigned. In particular, we show that dative assignment during reanalysis triggers reaccess to the mental lexicon, giving rise to greater processing difficulty. We conclude with a discussion of empirical and theoretical consequences of our findings.


[Terahertz-band study on surface enhanced Raman scattering of nanoparticle].
Study on surface-enhanced Raman scattering in the terahertz-band proved in that the terahertz-band Raman enhancement also exists. By studing principles of electromagnetic enhancement of surface-enhanced Raman scattering, using the finite difference time-domain method, the electromagnetic enhancement of surface enhanced Raman scattering of nano-particles irradiated by terahertz-wave was simulated, and the enhancement effect of terahertz waves was analyzed. Simulation experiments show that using finite-difference time-domain method could obtain effectively accurate simulation result of nano-particle scattering, proving that for terahertz waves, surface-enhanced effects on the surface of the nano-particle also exist. The results for surface enhanced Raman scattering extended from the visible and infrared to terahertz-band, and provide a basis for application of the combination of surface-enhanced Raman scattering and terahertz-wave.


Ultrasound-guided permanent implantation of peripheral nerve stimulation (PNS) system for neuropathic pain of the extremities: original cases and outcomes.
Our objective is to describe our experience applying a minimally invasive, ultrasound-assisted technique for peripheral nerve stimulation (PNS) trial and permanent implantation in patients with neuropathic extremity pain. Retrospective case series and review of the literature. Setting. Tertiary referral center and academic medical center. Patients with upper or lower extremity neuropathic pain resistant to other therapies who responded to an ultrasound-guided peripheral nerve block at a proximal location. Interventions. Ultrasound-assisted through-the-needle placement of percutaneous neurostimulation electrodes on target major peripheral nerves for fully percutaneous trial, staged trial or permanent implantation of PNS system. A PNS trial period of 3-7 days was used. Pain relief at last follow-up, complications, therapeutic limitations due to technique as applied. Results. Six of eight (75%) patients and 7/9 (78%) peripheral nerves had a successful trial and underwent permanent PNS system implantation using a minimally invasive, ultrasound-assisted technique from November 2007 to December 2008. All but one patient with an implanted PNS system had > or =50% pain relief at last follow-up and 3/7 (43%) permanent systems were associated with > or =80% relief. Loss of paresthesia required revision to dual-lead systems in upper extremity radial nerve PNS. Infection led to explant in one case. In a small series of patients, a minimally invasive, ultrasound-assisted technique for PNS trial, and permanent PNS implantation proved feasible. Patients without adequate analgesia during neurostimulation trial avoided surgical incision and those undergoing permanent implantation were not subjected to the potential morbidity associated with nerve dissection.


Norwogonin flavone suppresses the growth of human colon cancer cells via mitochondrial mediated apoptosis, autophagy induction and triggering G2/M phase cell cycle arrest.
Colorectal cancer is one of the deadly malignancies and is one of the top three most common cancers and the third leading cause of cancer-related deaths. The main objective of the study was to investigate the anticancer effects of norwogonin - a naturally occurring plant flavone. We also examined its effects on programmed cell death, autophagy and cell cycle phase distribution. Cell viability of colon cancer cells was evaluated by MTT assay while apoptotic studies were carried out by fluorescence microscopy using acridine orange (AO)/ethidium bromide (EB) and Comet assays. Transmission electron microscopy (TEM) was used to study formation of autophagosomes reminiscent of autophagy. Furthermore, western blot assay was used to study the effects of norwogonin on apoptosis-related protein expressions including Bax, Bcl-2 and autophagy-related proteins. Effects on cell cycle were evaluated by flow cytometry. The results showed that norwogonin causes substantial reduction in the viability of the human colorectal carcinoma cells in a dose-dependent manner, exhibiting an IC50 of 15.5 µM in cancer cells and IC50 of 90 µM in normal cell lines. The AO/EB staining assay showed that norwogonin suppresses the viability of cancer cells via induction of apoptotic cell death which was associated with increase in Bax and decrease in Bcl-2 levels. Comet assay results also confirmed that norwogonin induces apoptosis. Norwogonin also led to induction of autophagy along with triggering G2/M phase cell cycle arrest. In conclusion, the current study shows that norwogonin has a potential to inhibit in vitro colorectal cancer cells growth by triggering apoptosis, autophagy and cell cycle arrest and as such could be developed as a possible anticancer agent.


Combined approach for embolization of otherwise unmanageable gastric varices.
This study aimed to determine the feasibility, safety and effectiveness of combined percutaneous transhepatic obliteration (PTO) and balloon-occluded retrograde transvenous obliteration (BRTO) therapy for the treatment of patients with high-risk bleeding gastric varices. Ten patients were retrospectively reviewed. All the patients presented gastric varices, according to the Sarin classification, at high risk of bleeding, and not otherwise manageable. Patients with portal vein thrombosis were excluded. All patients were treated with a combination of PTO and BRTO. In all cases the gastric varices were embolized with glue, combined with coils or not, with an occlusion balloon inflated into the shunt. In 7 cases, embolization was immediate; in the remaining 3 the balloon remained inflated for 4 h and in 2 of them embolization of the shunt was required. Technical success was defined as complete obliteration of the gastric varices observed during a contrast-enhanced computed tomography study and endoscopy within 1 month following treatment. Clinical success was defined as absence of bleeding of gastric varices during the follow-up period. Major and minor complications during the follow up were recorded. Twelve sessions of combined PTO and BRTO procedures were performed in 10 patients; in 2 patients a new combined treatment was required during the follow up. Technical and clinical success was 100%. Neither major nor minor procedure-related complications were observed. Combined PTO and BRTO therapy is safe and effective for the treatment of gastric varices that cannot be managed otherwise.


Micropropagation of Prosopis chilensis (Mol.) Stuntz from young and mature plants.
Prosopis chilensis (Mol.) Stuntz (Algarrobo de Chile) is an important native tree species that can be grown in arid and semiarid regions for wood and forage production and environmental protection. Developing a simple and reliable in vitro protocol for cloning it would enable to improve it genetically. Explants of P. chilensis were taken from 4 months-old plants grown in the greenhouse or from adult trees grown in a natural environment. Nodal segments 1-2 cm long containing an axillary bud were selected from elongating shoots. These cuttings were aseptically cultured on two agar-solid basal media, MS or BTMm, and treated with 0.05 mg L-1 BA and 3 mg L-1 of either IAA, IBA or NAA. Sucrose (3% w/v) was used as carbon source. The percentage of sprouted cuttings and whole plant regeneration as well as its shoot and root length were recorded. Number, length and dry weight of shoots and roots were also measured. Rooting was successful with cuttings taken from young or adult plants, but explants from young plants showed a better response. Culturing in BTMm resulted in significantly greater shoot and root biomass than culturing in MS. Moreover, this response was higher in young explants when IBA was used as growth regulator. This paper reports a simple and effective method to micropropagate P. chilensis from young and adult plants.


Evolution of a histone H4-K16 acetyl-specific DNA aptamer.
We report the in vitro selection of DNA aptamers that bind to histone H4 proteins acetylated at lysine 16. The best aptamer identified in this selection binds to the target protein with a K(d) of 21 nM and discriminates against both the nonacetylated protein and histone H4 proteins acetylated at lysine 8. Comparative binding assays performed with a chip-quality antibody reveal that this aptamer binds to the acetylated histone target with similar affinity to a commercial antibody but shows significantly greater specificity (15-fold versus 2400-fold) for the target molecule. This result demonstrates that aptamers that are both modification and location specific can be generated to bind specific protein post-translational modifications.


Effects of salicylates and aminoglycosides on spontaneous otoacoustic emissions in the Tokay gecko.
The high sensitivity and sharp frequency discrimination of hearing depend on mechanical amplification in the cochlea. To explore the basis of this active process, we examined the pharmacological sensitivity of spontaneous otoacoustic emissions (SOAEs) in a lizard, the Tokay gecko. In a quiet environment, each ear produced a complex but stable pattern of emissions. These SOAEs were reversibly modulated by drugs that affect mammalian otoacoustic emissions, the salicylates and the aminoglycoside antibiotics. The effect of a single i.p. injection of sodium salicylate depended on the initial power of the emissions: ears with strong control SOAEs displayed suppression at all frequencies, whereas those with weak control emissions showed enhancement. Repeated oral administration of acetylsalicylic acid reduced all emissions. Single i.p. doses of gentamicin or kanamycin suppressed SOAEs below 2.6 kHz, while modulating those above 2.6 kHz in either of two ways. For ears whose emission power at 2.6-5.2 kHz encompassed more than half of the total, individual emissions displayed facilitation as great as 35-fold. For the remaining ears, emissions dropped to as little as one-sixth of their initial values. The similarity of the responses of reptilian and mammalian cochleas to pharmacological intervention provides further evidence for a common mechanism of cochlear amplification.


Comparison of side chain interactions performed by structurally equivalent residues in homologous protein structures.
The present work describes the computer program Hom-Bond, which allows to identify and compare intra-molecular interactions performed by side chain polar atoms as observed in a family of homologous protein structures with known and conserved 3-D conformation. For this purpose, the side chain to side chain and the side chain to main chain hydrogen bonds, the disulfide and the salt bridges are identified in each considered protein structure. Subsequently, the side chain interactions are displayed according to the multiple sequence alignment. The presented approach allows to easily identify bonds which are conserved in homologous proteins and to analyse rearrangements of the network of side chain interactions that characterize each protein structure.


The Contribution of Adolescent Effortful Control to Early Adult Educational Attainment.
Effortful control has been proposed as a set of neurocognitive competencies that is relevant to self-regulation and educational attainment (Posner & Rothbart, 2007). This study tested the hypothesis that a multiagent report of adolescents' effortful control (age 17) would be predictive of academic persistence and educational attainment (age 23-25), after controlling for other established predictors (family factors, problem behavior, grade point average, and substance use). Participants were 997 students recruited in 6th grade from 3 urban public middle schools (53% males; 42.4% European American; 29.2% African American). Consistent with the hypothesis, the unique association of effortful control with future educational attainment was comparable in strength to that of parental education and students' past grade point average, suggesting that effortful control contributes to this outcome above and beyond well-established predictors. Path coefficients were equivalent across gender and ethnicity (European Americans and African Americans). Effortful control appears to be a core feature of the self-regulatory competencies associated with achievement of educational success in early adulthood. These findings suggest that the promotion of self-regulation in general and effortful control in particular may be an important focus not only for resilience to stress and avoidance of problem behavior, but also for growth in academic competence.


Nanoparticle penetration and transport in living pumpkin plants: in situ subcellular identification.
In recent years, the application of nanotechnology in several fields of bioscience and biomedicine has been studied. The use of nanoparticles for the targeted delivery of substances has been given special attention and is of particular interest in the treatment of plant diseases. In this work both the penetration and the movement of iron-carbon nanoparticles in plant cells have been analyzed in living plants of Cucurbita pepo. The nanoparticles were applied in planta using two different application methods, injection and spraying, and magnets were used to retain the particles in movement in specific areas of the plant. The main experimental approach, using correlative light and electron microscopy provided evidence of intracellular localization of nanoparticles and their displacement from the application point. Long range movement of the particles through the plant body was also detected, particles having been found near the magnets used to immobilize and concentrate them. Furthermore, cell response to the nanoparticle presence was detected. Nanoparticles were capable of penetrating living plant tissues and migrating to different regions of the plant, although movements over short distances seemed to be favoured. These findings show that the use of carbon coated magnetic particles for directed delivery of substances into plant cells is a feasible application.


Inhibition of host protein synthesis by Sindbis virus: correlation with viral RNA replication and release of nuclear proteins to the cytoplasm.
Infection of mammalian cells by Sindbis virus (SINV) profoundly blocks cellular mRNA translation. Experimental evidence points to viral non-structural proteins (nsPs), in particular nsP2, as the mediator of this inhibition. However, individual expression of nsP1, nsP2, nsP3 or nsP1-4 does not block cellular protein synthesis in BHK cells. Trans-complementation of a defective SINV replicon lacking most of the coding region for nsPs by the co-expression of nsP1-4 propitiates viral RNA replication at low levels, and inhibition of cellular translation is not observed. Exit of nuclear proteins including T-cell intracellular antigen and polypyrimidine tract-binding protein is clearly detected in SINV-infected cells, but not upon the expression of nsPs, even when the defective replicon was complemented. Analysis of a SINV variant with a point mutation in nsP2, exhibiting defects in the shut-off of host protein synthesis, indicates that both viral RNA replication and the release of nuclear proteins to the cytoplasm are greatly inhibited. Furthermore, nucleoside analogues that inhibit cellular and viral RNA synthesis impede the blockade of host mRNA translation, in addition to the release of nuclear proteins. Prevention of the shut-off of host mRNA translation by nucleoside analogues is not due to the inhibition of eIF2α phosphorylation, as this prevention is also observed in PKR(-/-) mouse embryonic fibroblasts that do not phosphorylate eIF2α after SINV infection. Collectively, our observations are consistent with the concept that for the inhibition of cellular protein synthesis to occur, viral RNA replication must take place at control levels, leading to the release of nuclear proteins to the cytoplasm.


Overexpression of MLPH in Rectal Cancer Patients Correlates with a Poorer Response to Preoperative Chemoradiotherapy and Reduced Patient Survival.
Data mining of a public transcriptomic rectal cancer dataset (GSE35452) from the Gene Expression Omnibus, National Center for Biotechnology Information identified the melanophilin (MLPH) gene as the most significant intracellular protein transport-related gene (GO:0006886) associated with a poor response to preoperative chemoradiation. An MLPH immunostain was performed on biopsy specimens from 172 rectal cancer patients receiving preoperative chemoradiation; samples were divided into high- and low-expression groups by H-scores. Subsequently, the correlations between MLPH expression and clinicopathologic features, tumor regression grade, disease-specific survival (DSS), local recurrence-free survival (LRFS), and metastasis-free survival (MeFS) were analyzed. MLPH expression was significantly associated with CEA level (p = 0.001), pre-treatment tumor status (p = 0.022), post-treatment tumor status (p < 0.001), post-treatment nodal status (p < 0.001), vascular invasion (p = 0.028), and tumor regression grade (p < 0.001). After uni- and multi-variable analysis of five-year survival, MLPH expression was still associated with lower DSS (hazard ratio (HR), 10.110; 95% confidence interval (CI), 2.178-46.920; p = 0.003) and MeFS (HR, 5.621; 95% CI, 1.762-17.931; p = 0.004). In conclusion, identifying MLPH expression could help to predict the response to chemoradiation and survival, and aid in personal therapeutic modification.


X-ray-Powered Micromotors.
Light-powered wireless manipulation of small objects in fluids has been of interest for biomedical and environmental applications. Although many techniques employing UV-vis-NIR light sources have been devised, new methods that hold greater penetrating power deep into medium are still in demand. Here, we develop a method to exploit X-rays to propel half-metal-coated Janus microparticles in aqueous solution. The Janus particles are simultaneously propelled and visualized in real-time by using a full-field transmission X-ray microscope. Our real-time observation discovers that the propulsive motion follows the bubble growth enhanced by water radiolysis near the particle surface under X-ray irradiation. We also show that the propulsion speed is remotely controlled by varying the radiation dose. We expect this work to open opportunities to employ light-powered micro/nanomotors in opaque environments, potentially by combining with medical imaging or nondestructive testing.


[The basics on mechanical ventilation support in acute respiratory distress syndrome].
Acute Respiratory Distress Syndrome (ARDS) is understood as an inflammation-induced disruption of the alveolar endothelial-epithelial barrier that results in increased permeability and surfactant dysfunction followed by alveolar flooding and collapse. ARDS management relies on mechanical ventilation. The current challenge is to determine the optimal ventilatory strategies that minimize ventilator-induced lung injury (VILI) while providing a reasonable gas exchange. The data support that a tidal volume between 6-8 ml/kg of predicted body weight providing a plateau pressure < 30 cmH₂O should be used. High positive end expiratory pressure (PEEP) has not reduced mortality, nevertheless secondary endpoints are improved. The rationale used for high PEEP argues that it prevents cyclic opening and closing of airspaces, probably the major culprit of development of VILI. Chest computed tomography has contributed to our understanding of anatomic-functional distribution patterns in ARDS. Electric impedance tomography is a technique that is radiation-free, but still under development, that allows dynamic monitoring of ventilation distribution at bedside.


Cucurbit[n]uril analogues: synthetic and mechanistic studies.
[reaction: see text] The synthesis of cucurbit[n]uril analogues (18, 19, (+/-)-20, 33, 34, 35, 36, and 37) is presented. These CB[5], CB[6], and CB[7] analogues all contain bis(phthalhydrazide) walls that are incorporated into the macrocycle. The tailor-made synthesis of these CB[n] analogues proceeds by the condensation of the appropriate bis(electrophile) (4, 7, or 9) with bis(phthalhydrazide) (17), which delivers the CB[6] and CB[7] analogues in good yield, whereas the CB[5] analogue is formed in low yield. To improve the solubility characteristics of the CB[n] analogues for recognition studies in water or organic solution, the CO2Et groups were transformed to CO2H and CO2(CH2)9CH3 groups. On the basis of the results of product resubmission experiments, we conclude that these macrocycles are kinetic products. To help rationalize the good yields obtained in the CB[6] and CB[7] analogue macrocyclization reactions, we performed mechanistic studies of model methylene bridged glycoluril dimers, which suggest an intramolecular isomerization during CB[n] analogue formation.


Acquired hematopoietic stem cell defects determine B-cell repertoire changes associated with aging.
Aging is associated with an inability to mount protective antibody responses to vaccines and infectious agents. This decline is associated with acquisition of defects in multiple cellular compartments, including B cells. While peripheral B-cell numbers do not decline with aging, the composition of the compartment appears to change, with loss of naïve follicular B cells, accumulation of antigen-experienced cells, and alteration of the antibody repertoire. The underlying cause of this change is unknown. We tested the hypothesis that aging-associated repertoire changes can be attributed directly to decreased B lymphopoiesis. Using an Ig transgenic model to report changes in the B-cell repertoire, we show that the reduced B-cell generative capacity of "aged" long-term reconstituting hematopoietic stem cells (LT-HSCs) alters the representation of antigen specificities in the peripheral B-cell repertoire. Further, we show that reconstitution using suboptimal numbers of fully functional LT-HSCs results in the generation of a similarly altered B-cell repertoire. This may be an important factor to consider when deciding the number of bone marrow cells to transplant in the clinical setting. In conclusion, when B lymphopoiesis is limited peripheral B-cell homeostasis is altered. This is reflected in reduced diversity of the B-cell repertoire, which likely reduces the protective quality of the immune response.


Human bocavirus infection in Belgian children with respiratory tract disease.
Human bocavirus (HBoV) has been detected primarily in children with acute lower respiratory tract disease (LRTD), but its occurrence, clinical profile, and role as a causative agent of RTD are not clear. The aim of this study was to investigate the prevalence and the potential clinical relevance of HBoV. Using molecular tests, we tested 1352 nasopharyngeal samples obtained between October 1, 2017 and April 30, 2018 from children up to the age of 16 with RTD for the presence of HBoV DNA and 20 other respiratory pathogens at three different hospitals in Belgium. HBoV was detected in 77 children with a median age of 10.6 months. Consecutive samples were available for 15 HBoV-positive children and showed persistent HBoV positivity in four of them. Monoinfection was observed in six infants. Four of them were born prematurely and were infected during hospitalization at the neonatal intensive care unit (NICU). Only one of these six monoinfected children was diagnosed with recurrent wheezing due to HBoV. This child was carried to term and had a high viral load. Coinfections, most frequently with rhinovirus (52.1%) and adenovirus (49.3%), were observed in 72 patients. In seventeen of them in which HBoV was present at high viral load or higher viral load than its copathogens, bronchi(oli)tis (n = 8), recurrent wheezing (n = 8) or episodic wheezing (n = 1) were diagnosed. Our results suggest that HBoV infection at high viral load in infants is associated with wheezing (P = 0.013, Cramer's V = 0.613).


Severe COVID-19 Pneumonia in a 30-Year-Old Woman in the 36th Week of Pregnancy Treated with Postpartum Extracorporeal Membrane Oxygenation.
BACKGROUND There are few reports of coronavirus disease 2019 (COVID-19) in pregnant women. Although coagulation dysfunction was reported to affect the severity of COVID-19, the association between pregnancy, which is usually accompanied by changes in coagulation function, and the worsening of COVID-19 is unknown. We present a case of a 30-year-old woman in the 36th week of pregnancy who was diagnosed with severe COVID-19 pneumonia and required postpartum extracorporeal membrane oxygenation (ECMO) therapy. CASE REPORT A 30-year-old, 36-weeks pregnant woman presented to our hospital and was diagnosed with severe COVID-19 pneumonia soon after she had undergone a cesarean section. Her respiratory failure could not be managed by conventional therapeutic approaches. Therefore, ECMO was administered on day 7. Controlling coagulation function to maintain ECMO therapy was challenging. Nafamostat mesylate and cryoprecipitate were administered to treat the hypercoagulative status and severe hypofibrinogenemia, respectively. Since coagulopathy and her respiratory state improved, the ECMO therapy was terminated on day 15. CONCLUSIONS We report a case of severe COVID-19 pneumonia in a pregnant woman urgently treated with ECMO in the postpartum period. Thus, this case highlights the importance of close monitoring and appropriate medical care for pregnant women with severe COVID-19 pneumonia.


Characterization of Spontaneous Vascular Findings in the Papillary Muscles of Beagle Dogs.
Standard histology trimming practices for toxicity studies in dogs include preparation of a section of the papillary muscle associated with the left ventricular free wall-the anteriolateral or anterior muscle. In contrast, the posteromedial or posterior papillary muscle, associated with the interventricular septum, is not commonly evaluated. In humans, the posterior papillary muscle is more often affected in ischemic myocardial disease, in large part due to the absence of collateral circulation, in contrast to the anterior muscle. Due to the differential vascular supply to the papillary muscles, we sought to determine whether there is a higher incidence of spontaneous coronary vascular changes in the posterior papillary muscle versus the anterior muscle in dogs. The hearts of 30 vehicle-treated or untreated beagle dogs that were euthanized for other purposes were collected and sectioned in a consistent manner to include both papillary muscles for histologic evaluation. The posterior muscle consistently had higher incidences of intramural coronary arteries affected by vascular medial hypertrophy, medial arteriosclerosis, intimal hyperplasia, and/or disruption or loss of the internal elastic lamina. This observation may have significant implications for the evaluation and characterization of spontaneous and xenobiotic-induced cardiovascular lesions in dogs.


Aging and insulin secretion.
Glucose tolerance progressively declines with age, and there is a high prevalence of type 2 diabetes and postchallenge hyperglycemia in the older population. Age-related glucose intolerance in humans is often accompanied by insulin resistance, but circulating insulin levels are similar to those of younger people. Under some conditions of hyperglycemic challenge, insulin levels are lower in older people, suggesting beta-cell dysfunction. When insulin sensitivity is controlled for, insulin secretory defects have been consistently demonstrated in aging humans. In addition, beta-cell sensitivity to incretin hormones may be decreased with advancing age. Impaired beta-cell compensation to age-related insulin resistance may predispose older people to develop postchallenge hyperglycemia and type 2 diabetes. An improved understanding of the metabolic alterations associated with aging is essential for the development of preventive and therapeutic interventions in this population at high risk for glucose intolerance.


Care related pain and functional restoration for chronic low back pain: a prospective study.
To assess care related pain during a program of functional restoration for chronic low back pain. Chronic low back pain patients were prospectively included in a program of functional restoration. Low back pain was recorded before and after each morning and afternoon sessions, all over the program of functional restoration, for equivalence study. Equivalence was admitted if 95% confidence interval of the difference in pain falled wholly in the interval ± 10/100mm. Medication was recorded during the program. Progression of the patients during the program and 3-month effectiveness were also recorded for internal validity. Thirty-seven patients were included. They underwent a progression in load and spent energy during the program. Follow-up also indicated improvement of functional ability and quality of life. There was no variation in pain during the program however medication intake increased. There was no significant care related pain during our program of functional restoration for chronic low back pain. This may be due to adaptation of the medication and suggests the importance of close medical management of patients during functional restoration.


Confocal pore size measurement based on super-resolution image restoration.
A confocal pore size measurement based on super-resolution image restoration is proposed to obtain a fast and accurate measurement for submicrometer pore size of nuclear track-etched membranes (NTEMs). This method facilitates the online inspection of the pore size evolution during etching. Combining confocal microscopy with super-resolution image restoration significantly improves the lateral resolution of the NTEM image, yields a reasonable circle edge-setting criterion of 0.2408, and achieves precise pore edge detection. Theoretical analysis shows that the minimum measuring diameter can reach 0.19 μm, and the root mean square of the residuals is only 1.4 nm. Edge response simulation and experiment reveal that the edge response of the proposed method is better than 80 nm. The NTEM pore size measurement results obtained by the proposed method agree well with that obtained by scanning electron microscopy.


Quantitative assessment of changes in carotid plaques during cilostazol administration using three-dimensional ultrasonography and non-gated magnetic resonance plaque imaging.
Cilostazol, an antiplatelet agent, is reported to induce the regression of atherosclerotic changes. However, its effects on carotid plaques are unknown. Hence, we quantitatively investigated the changes that occur within carotid plaques during cilostazol administration using three-dimensional (3D) ultrasonography (US) and non-gated magnetic resonance (MR) plaque imaging. We prospectively examined 16 consecutive patients with carotid stenosis. 3D-US and T1-weighted MR plaque imaging were performed at baseline and 6 months after initiating cilostazol therapy (200 mg/day). We measured the volume and grayscale median (GSM) of the plaques from 3D-US data. We also calculated the contrast ratio (CR) of the carotid plaque against the adjacent muscle and areas of the intraplaque components: fibrous tissue, lipid, and hemorrhage components. The plaque volume on US decreased significantly (median at baseline and 6 months, 0.23 and 0.21 cm(3), respectively; p = 0.03). In the group exhibiting a plaque volume reduction of more than 10%, GSM on US increased significantly (24.8 and 71.5, respectively; p = 0.04) and CR on MRI decreased significantly (1.13 and 1.04, respectively; p = 0.02). In this group, in addition, the percent area of the fibrous component on MRI increased significantly (68.6% and 79.4%, respectively; p = 0.02), while those of the lipid and hemorrhagic components decreased (24.9% and 20.5%, respectively; p = 0.12) (1.0% and 0.0%, respectively; p = 0.04). There were no substantial changes in intraplaque characteristics in either US or MRI in the other group. 3D-US and MR plaque imaging can quantitatively detect changes in the size and composition of carotid plaques during cilostazol therapy.


Carbonation of steel slag for CO2 sequestration: leaching of products and reaction mechanisms.
Carbonation of industrial alkaline residues can be used as a CO2 sequestration technology to reduce carbon dioxide emissions. In this study, steel slag samples were carbonated to a varying extent. Leaching experiments and geochemical modeling were used to identify solubility-controlling processes of major and trace elements, both with regard to carbonation mechanisms and the environmental properties of the (carbonated) steel slag. Carbonation was shown to reduce the leaching of alkaline earth metals (except Mg) by conversion of Ca-phases, such as portlandite, ettringite, and Ca-(Fe)-silicates into calcite, possibly containing traces of Ba and Sr. The leaching of vanadium increased substantially upon carbonation, probably due to the dissolution of a Ca-vanadate. The reactive surface area of Al- and Fe-(hydr)oxides increased with the carbonation degree, which tends to reduce the leaching of sorption-controlled trace elements. Sorption on Mn- (hydr)oxides was found to be required to adequately model the leaching of divalent cations, but was not influenced by carbonation. Consideration of these three distinct reactive surfaces and possible (surface) precipitation reactions resulted in adequate modeling predictions of oxyanion and trace metal leaching from (carbonated) steel slag. Hence, these surfaces exert a major influence on the environmental properties of both fresh and carbonated steel slag.


Morbidity associated with gastrostomy placement in children demands an ongoing integrated approach to care.
To evaluate the short and long term morbidity of gastrostomy insertion, and to identify ongoing management requirements. A retrospective review was undertaken of the hospital casenotes of children aged up to fifteen years who had a gastrostomy placed in Christchurch over a six year period to March 1998. 42 children had a gastrostomy fashioned, 35 in the last three years of the period reviewed. The most common underlying diagnosis was neurological disease (48%), and the most common indication for tube placement was failure to feed orally. Complications were frequent but minor. Morbidity was related to local erythema and infection around the stoma (85 episodes in 23 children), persistent and major gastric fluid leakage (three episodes), and mechanical failure of the tube (21 episodes). Gastro-oesophageal reflux was seen in fourteen children, nine of whom had primary neurological disease. Complications were seen more after open gastrostomy than after percutaneous endoscopic placement (6.6:4.7). There was no mortality related directly to the gastrostomy tube or tube placement. An increase in the frequency of gastrostomy placements has been seen over this period. As the number of children with a gastrostomy increases, so too have the demands on medical and nursing staff to care for and manage the devices. The frequency of minor ongoing problems necessitates ongoing support of the child and care of the gastrostomy. A close working relationship between outreach nursing staff, stoma therapists and medical staff is required if morbidity is to be minimised. Education, audit and review remain important additional aspects of care.


Normal ovaries in neonates and infants: a sonographic study of 77 patients 1 day to 24 months old.
Normal values for ovarian measurements in adults have been revised over the past decade. A recent report stated that ovarian cysts were common in healthy girls 2-13 years old, refuting the findings of a 1984 study. No large sonographic study of normal ovaries in girls 1 day to 24 months old has been performed. We evaluated ovaries in girls in this age group to determine the normal volume and prevalence of ovarian cysts. The ovaries of 77 consecutive patients 1 day to 24 months old were evaluated during routine pelvic sonography. Patients were divided into three age groups: 1 day to 3 months old (when gonadotropin levels are highest because of loss of placental hormonal influence), 4-12 months old (an intermediate group), and 13-24 months old (when gonadotropin levels are low). Ninety-eight ovaries were imaged in three dimensions. The mean volume was 1.06 cm3 (range, 0.7-3.6 cm3) among girls up to 3 months old; 1.05 cm3 (range, 0.2-2.7 cm3) among girls 4-12 months old; and 0.67 cm3 (range, 0.1-1.7 cm3) among girls 13-24 months old. We found no significant difference in mean volumes among the three groups. The prevalence of ovarian cysts was similar in all three groups; ovarian cysts were seen in 84% of all imaged ovaries. Macrocysts (cysts larger than 9 mm) were seen in 18% of all cystic ovaries. Ovaries of girls 1 day to 24 months old can have volumes greater than 1 cm3. Ovarian cysts are common. Macrocysts can be seen despite claims that they are rare in girls less than 11 years old.


Advanced oxidation process based on the Cr(III)/Cr(VI) redox cycle.
Oxidative degradation of aqueous organic pollutants, using 4-chlorophenol (4-CP) as a main model substrate, was achieved with the concurrent H(2)O(2)-mediated transformation of Cr(III) to Cr(VI). The Fenton-like oxidation of 4-CP is initiated by the reaction between the aquo-complex of Cr(III) and H(2)O(2), which generates HO(•) along with the stepwise oxidation of Cr(III) to Cr(VI). The Cr(III)/H(2)O(2) system is inactive in acidic condition, but exhibits maximum oxidative capacity at neutral and near-alkaline pH. Since we previously reported that Cr(VI) can also activate H(2)O(2) to efficiently generate HO(•), the dual role of H(2)O(2) as an oxidant of Cr(III) and a reductant of Cr(VI) can be utilized to establish a redox cycle of Cr(III)-Cr(VI)-Cr(III). As a result, HO(•) can be generated using both Cr(III)/H(2)O(2) and Cr(VI)/H(2)O(2) reactions, either concurrently or sequentially. The formation of HO(•) was confirmed by monitoring the production of p-hydroxybenzoic acid from [benzoic acid + HO(•)] as a probe reaction and by quenching the degradation of 4-CP in the presence of methanol as a HO(•) scavenger. The oxidation rate of 4-CP in the Cr(III)/H(2)O(2) solution was highly influenced by pH, which is ascribed to the hydrolysis of Cr(III)(H(2)O)(n) into Cr(III)(H(2)O)(n-m)(OH)(m) and the subsequent condensation to oligomers. The present study proposes that the Cr(III)/H(2)O(2) combined with Cr(VI)/H(2)O(2) process is a viable advanced oxidation process that operates over a wide pH range using the reusable redox cycle of Cr(III) and Cr(VI).


Current Situation of the Presence of Dirofilaria immitis in Dogs and Humans in Bucaramanga, Colombia.
The cardiopulmonary dirofilariosis caused by Dirofilaria immitis, is a vector-borne infection, which can be transmitted to humans. The main hosts are both domestic and wild canids. This species mainly occurs in tropical and subtropical climates, and temperature and humidity are the main factors that favor the presence and proliferation of culicid mosquitoes as vectors of the disease. There are few reports of cardiopulmonary dirofilariosis in dogs and humans in Colombia, a region with favorable climatic conditions which favors the presence of mosquitoes that act as vectors of the disease. Therefore, this study aimed to examine its current prevalence in dogs and the risk of human exposure to the disease in Bucaramanga, one of the most populated areas in Colombia located at the center of the country. Furthermore, its demographic and environmental characteristics could be useful as a study model for other similar locations and neighboring countries. Serum samples from 351 dogs and 506 humans from the Bucaramanga Metropolitan area were analyzed. All dog samples were analyzed by Knott's technique and tested with a commercial immunochromatographic to detect the presence of circulating antigens of D. immitis. Human samples were analyzed using a non-commercial ELISA test kit to detect IgG against the somatic antigens of adult D. immitis and Wolbachia. Positive results were further confirmed using western blot analysis. Thirty-eight dogs tested positive with a overall prevalence of 10.82%. Of these dogs, 18 showed D. immitis microfilariae, being 5.12% of the total population. The overall seroprevalence in humans was 6.71%; seroprevalence was significantly higher in individuals aged 16-34 years-old and in women than in men. To our knowledge, this study describes seropositivity to D. immitis for the first time in a Colombian human population located in the same area as that of dogs infected with D. immitis, which represents a potential threat to public health. In humans, age and gender can be considered risk factors for exposure to D. immitis.


Demonstration of reversible complete left bundle branch block following tachycardia-induced cardiomyopathy.
A 56-year-old man with atrial fibrillation and complete left bundle branch block (CLBBB) developed heart failure refractory to the initial medical treatment. Both the CLBBB and cardiac dysfunction completely recovered only with an advanced medical regimen for rate control and heart failure. This report describes a case with reversible CLBBB following tachycardia-induced cardiomyopathy, who was not a candidate for cardiac resynchronization therapy.


Combustion Products of Calcium Carbide Reused by Cu-Based Catalysts for Acetylene Carbonylation.
Sustainable development is a worldwide concern. This work mainly focuses on the reuse of the combustion products of calcium carbide and the influence of different kinds of copper on the acetylene carbonylation reaction. A series of catalysts were prepared by heating the precursors under various atmospheres (air, hydrogen, and nitrogen). The X-ray diffraction and the X-ray photoelectron spectroscopy have been analyzed regarding copper species composition and content in catalysts. The result of the Cu+-promoted reaction was in good agreement with the conducted density functional theory analysis, and we speculate that Cu+ promotes the transfer of electrons in the reaction. Transmission electron microscopy and elemental mapping evaluation confirmed the difference in Cu dispersion. Characterization of catalysts using temperature programmed desorption and pyridine Fourier-transform infrared revealed differences in their acidity. Acidity was found to be favorable for acetylene carbonylation. Selectivity and yield of the CuAlZn-LDO(N) catalyst at 225 °C were 73 and 70%, respectively, and the catalyst showed good stability over two consecutive cycles of reuse.


Verification of the position of a central venous catheter by intra-atrial ECG. When does this method fail?
The purpose of this study was to determine why intra-atrial ECG tracing for checking the position of a central venous catheter fails in certain patients. Three hundred and fifty prospective and consecutive patients scheduled for central venous catheterization using various puncture sites and techniques were investigated. The catheters were 20 cm in length. After its introduction, the catheter was connected to an Alphacard (Sterimed, Saarbrücken) for the intra-atrial ECG tracing. The method failed in 29 patients, of whom nine had manifest myocardial pathology. In two patients the catheter looped, while in the remaining 18 the catheter proved to be too short. In these 18 patients, the cannulation was mainly performed via the external jugular vein and/or from the left side. Most of the patients were elderly males, and 11 of the 18 patients showed radiological signs of pulmonary emphysema. In such individuals it is advisable to use a catheter longer than 20 cm.


A filarial nematode-secreted phosphorylcholine-containing glycoprotein uncouples the B cell antigen receptor from extracellular signal-regulated kinase-mitogen-activated protein kinase by promoting the surface Ig-mediated recruitment of Src homology 2 domain-containing tyrosine phosphatase-1 and Pac-1 mitogen-activated kinase-phosphatase.
Unraveling the molecular mechanisms by which filarial nematodes, major human pathogens in the tropics, evade the host immune system remains an elusive goal. We have previously shown that excretory-secretory product-62 (ES-62), a homologue of phosphorylcholine-containing molecules that are secreted by human parasites and which is active in rodent models of filarial infection, is able to polyclonally activate certain protein tyrosine kinase and mitogen-activating protein kinase signal transduction elements in B lymphocytes. Such activation mediates desensitization of subsequent B cell Ag receptor (BCR) ligation-induced activation of extracellular signal-regulated kinase-mitogen-activated protein (ErkMAP) kinase and ultimately B cell proliferation. We now show that the desensitization is due to ES-62 targeting two major regulatory sites of B cell activation. Firstly, pre-exposure to ES-62 primes subsequent BCR-mediated recruitment of SHP-1 tyrosine phosphatase to abolish recruitment of the RasErkMAP kinase cascade via the Igalphabeta-ShcGrb2Sos adaptor complex interactions. Secondly, any ongoing ErkMAP kinase signaling in ES-62-primed B cells is terminated by the MAP kinase phosphatase, Pac-1 that is activated consequently to challenge via the BCR.


Assessment of an evidence-based laryngeal cancer fact sheet: A mixed methods study.
To evaluate perceptions of a laryngeal cancer fact sheet amongst people with direct experience of the disease and its treatment. A mixed methods study (questionnaire and interview) evaluating the information resource was conducted across two institutions. In total 20 participants responded to the questionnaire. Overall participants reported the information resource was detailed and understandable. Insufficient information was provided on: impact on family in eight participants (40%); impact on work in six (33%); and, second opinions and long-term side effects in five (25%). The majority (67%) wanted a large amount of information with the preferred source being one-on-one meetings with their doctor. The thematic analysis identified three main themes: preferences for information, self-management; and, information sources. People with direct experience of laryngeal cancer and its treatments reported the information resource was comprehensive and clear. There were some gaps in the information provided, particularly related to survivorship issues.


[Intraoperative radiotherapy (IORT) for unresectable stage IVb pancreatic cancer].
We evaluated the efficacy of IORT for unresectable Stage IVb (Japan Pancreas Society classification) pancreatic cancer. Twelve patients were treated with IORT, 17 with external beam radiotherapy (ERT) and 17 with chemotherapy (CHT, 8 patients doxorubicin-based, 7 patients 5-FU-based). Survival, hospital-free survival and pain relief were compared among the three groups. In the IORT group, 7 patients underwent bypass surgery, 3 celiac plexus blockade, 3 ERT, 2 hyperthermia and 2 CHT. In the ERT group, 1 patient underwent bypass surgery, 7 hyperthermia and 14 CHT. Distant metastases were more frequently found in the CHT group than in the IORT group. Median survival and median hospital-free survival were 208 and 79 days in the IORT group, 125 and 32 days in the ERT group and 76 and 9 days in the CHT group, respectively. Pain relief was obtained in 45% (5/11) of symptomatic patients after IORT and in 27% (4/15) after ERT. No patient (0/13) in the CHT group experienced pain relief. In conclusion, our experience suggests that IORT can reduce pain and improve QOL in patients with unresectable pancreatic cancer.


Percutaneous extraction of a coiled, 20-year-old lead in a patient with cardiac resynchronization therapy.
A 61-year-old patient with a 20-year history of permanent pacemaker implantation and half--a-year cardiac resynchronization therapy using a left ventricular lead placed via surgical approach was admitted for extraction of an old coiled right ventricular lead, which triggered ventricular arrhythmia and created a risk of pulmonary embolism. The lead was extracted via the left femoral vein in two stages: untying a loop on the lead using a pig-tail catheter and Dotter basket followed by traction and dissection of adhesions using a Byrd dilator sheath. Dissection of the old lead from the active right ventricular one posed special technical problems.


EU-OPENSCREEN-chemical tools for the study of plant biology and resistance mechanisms.
EU-OPENSCREEN is an academic research infrastructure initiative in Europe for enabling researchers in all life sciences to take advantage of chemical biology approaches to their projects. In a collaborative effort of national networks in 16 European countries, EU-OPENSCREEN will develop novel chemical compounds with external users to address questions in, among other fields, systems and network biology (directed and selective perturbation of signalling pathways), structural biology (compound-target interactions at atomic resolution), pharmacology (early drug discovery and toxicology) and plant biology (response of wild or crop plants to environmental and agricultural substances). EU-OPENSCREEN supports all stages of a tool development project, including assay adaptation, high-throughput screening and chemical optimisation of the 'hit' compounds. All tool compounds and data will be made available to the scientific community. EU-OPENSCREEN integrates high-capacity screening platforms throughout Europe, which share a rationally selected compound collection comprising up to 300,000 (commercial and proprietary compounds collected from European chemists). By testing systematically this chemical collection in hundreds of assays originating from very different biological themes, the screening process generates enormous amounts of information about the biological activities of the substances and thereby steadily enriches our understanding of how and where they act.


Speckle Noise Algorithm-Based Ultrasound Imaging in Evaluating the Therapeutic Effect of Blood Purification on Children with Kidney Failure and Analysis of Its Correlation with Serum Inflammatory Factor Levels.
The study focused on the therapeutic effect of clinical treatment on urinary calculi with kidney failure and its relationship with the serum inflammatory factor levels. 90 children with melamine urinary calculi were selected as research subjects. Of them, 52 cases were in group 1 (nonrenal failure), and 38 cases were in group 2 (combined with renal failure). In addition, 35 hospitalized children with no history of melamine-contaminated milk feeding during the same period were used as healthy controls. They all underwent ultrasound imaging examination based on the speckle noise algorithm, and the prognosis was analyzed. It was found that the peak signal-to-noise ratio (PSNR), structural similarity (SSIM), and local edge preservation index (EPI) of the algorithm in this study were significantly greater than other algorithms (P < 0.05). The admission age of the children in group 1 was significantly younger than that of group 2, the bilateral stone rate was significantly higher than that in group 2, and the difference was statistically significant (P < 0.05). Of the 52 children in group 1, the stone disappeared in 25 cases after treatment, the stone was reduced in 20 cases, and the stone remained unchanged in 7 cases. The total effective rate of treatment was 88.46%. Of the 38 cases in group 2, the stone disappeared in 22 cases after treatment, the stone was reduced in 12 cases, and the stone remained unchanged in 4 cases. The total effective rate of treatment was 89.47%. No difference was noted in blood urea nitrogen (BUN), blood creatinine (Cr), TNF-α, and C-reactive protein (CRP) levels in group 1, group 2, and the healthy control group (P > 0.05). Hence, the denoising algorithm in this study has better denoising effects on ultrasound images than traditional algorithms, with higher definition and less noise and artifacts. The clinical treatment of children with urinary calculi and renal failure is highly effective, the renal function and serum inflammatory factor levels return to the normal range, and the inflammatory response is weakened.


New frontiers in the study of memory mechanisms.
We review recent work on three major lines of memory research: a) the possible role of the protein kinase M-zeta (PKMzeta) in memory persistence; b) the processes of "synaptic tagging and capture" in memory formation; c) the modulation of extinction learning, widely used in the psychotherapy of fear memories under the name of "exposure therapy". PKMzeta is a form of protein kinase C (PKC) that apparently remains stimulated for months after the consolidation of a given memory. Synaptic tagging is a mechanism whereby the weak activation of one synapse can tag it with a protein so other synapses in the same cell can reactivate it by producing other proteins that bind to the tag. Extinction, once mistakenly labeled as a form of forgetting, is by itself a form of learning; through it animals can learn to inhibit a response. We now know it can be modulated by neurotransmitters or by synaptic tagging, which should enable better control of its clinical use.


[Changes of arterial blood gas indexes of free-field primary blast lung injury of pigs and its application value].
To observe the changes of arterial blood gas indexes in pigs with the free-field primary blast lung injury (PBLI) model, and to explore the value of arterial blood gas indexes in predicting moderate to severe PBLI. Nine adult healthy Landrace pigs were selected to construct the pig free-field PBLI model. Arterial blood samples were taken 15 minutes before the explosion (before injury) and 10, 30, 60, 120, and 180 minutes after the explosion (after injury). Arterial blood gas indexes and pulse oxygen saturation (SpO2) were measured, compare the changes of blood gas analysis indexes and SpO2 levels at different time points, and observe the changes of gross injury scores and pathological injury scores of lung tissue. Analyze the correlation between the blood gas indicators. As time prolonged, at each time point, pH, arterial partial pressure of oxygen (PaO2), and SpO2 were lower than those before the injury, and blood lactic acid (Lac) and arterial partial pressure of carbon dioxide (PaCO2) were higher than those before the injury. Compared with that before the injury, the pH value in the blood decreased significantly 10 minutes after the injury (7.39±0.06 vs. 7.46±0.02, P < 0.05), and the Lac increased significantly (mmol/L: 3.61±2.89 vs. 1.10±0.28, P < 0.05), and lasts until 180 minutes after injury (pH value: 7.37±0.07 vs. 7.46±0.02, Lac (mmol/L): 2.40±0.79 vs. 1.10±0.28, both P < 0.05); while PaO2 and SpO2 decreased significantly at 180 minutes after injury [PaO2 (mmHg, 1 mmHg = 0.133 kPa): 59.40±10.94 vs. 74.81±9.39, P < 0.05; SpO2: 0.75±0.11 vs. 0.89±0.08, P < 0.05], PaCO2 increased significantly (mmHg: 56.17±5.38 vs. 48.42±4.93, P < 0.05). Correlation analysis showed that the gross injury score of lung blast injury animals was positively correlated with the pathological injury score (r = 0.866, P = 0.005); PaO2 and SpO2 were positively correlated (r = 0.703, P = 0.000); pH value and Lac were negative Correlation (r = -0.400, P = 0.006); pH value is negatively correlated with PaCO2 (r = -0.844, P = 0.000). This study successfully established a large mammalian free-field PBLI model, arterial blood gas analysis is helpful for the early diagnosis of PBLI, whether SpO2 can be used to evaluate the severity of lung injury remains to be further verified.


Raising a child with disability and dealing with life events: a mother's journey.
Intellectual disability services recognise people with intellectual disability as a family member and aim to provide effective support that facilitates family cohesion. However, the needs of families and individuals within the family should be addressed in a holistic manner to ensure a seamless service. This article outlines a mother's reflections on her experience of raising a child with autism spectrum disorder and its impact on her life. The article focuses on one mother's feelings, coping or lack of coping and growth during a time of sadness. This article written by a mother, nurse and academic through their shared interest in supporting families of children with a disability all came together to tell this story. It is hoped that this article will assist professionals understand a mother's perspective when raising a child with intellectual disability and support professionals to recognise the importance of listening, coordinated support and working in partnership.


Children with diabetes mellitus and family functioning: translating research into practice.
The specific challenges of living with diabetes mellitus (DM) which the child and parent must confront at each developmental stage are identified from infancy through adolescence. This discussion is divided into four developmental stages: 1) DM in infancy (0-2 years of age); 2) DM in toddlers and preschoolers (2-5 years); 3) DM in the school-age child (6-11 years); and 4) DM during early- to mid-adolescence (12-16 years). The central milestones of normal psychosocial development are reviewed and a summary is provided of recent research at each developmental stage with an emphasis on clinical implications for pediatric diabetes care teams.


Analysis of plasma distribution near the extraction region in surface produced negative ion sources.
In study of a negative ion source, it is important to understand the plasma characteristics near the extraction region. A recent experiment in the NIFS-R&D ion source has suggested that a "double ion plasma layer" which is a region consisting of hydrogen positive and negative ions exists near the plasma grid (PG). Density distribution of plasma near the extraction region is studied analytically. It is shown that the density distribution depends on an amount of the surface produced negative ions and the double ion plasma layer is formed near the PG surface for the case of strong surface production.


Comprehensive quality evaluation of Polygoni Orientalis Fructus and its processed product: chemical fingerprinting and simultaneous determination of seven major components coupled with chemometric analyses.
Polygoni Orientalis Fructus (POF) is a clinically effective Chinese medicine. Raw POF (RPOF) and POF Tostus (POFT) are used separately in clinics. However, incomplete progress has been made on quality control. To establish a comprehensive method for quality assessment of RPOF and POFT and to discriminate these two varieties. High-performance liquid chromatography combined with the diode array detector (HPLC-DAD) methods were developed for fingerprinting and quantitative analysis of seven major compounds in RPOF and POFT, and the main components were determined by HPLC-DAD coupled with Fourier-transform ion cyclotron resonance-mass spectrometry. Chemometric approaches were performed to discriminate RPOF and POFT and to screen discriminatory components. Fingerprints were established and 12 common peaks were identified, cannabisin G and cannabisin E were firstly identified from POF. In quantitative analysis, all analytes showed good regression (R > 0.9996) within test ranges and the recovery of the method was in the range 96.6-104.3%. Fingerprints in conjunction with similarity analysis and hierarchical clustering analysis (HCA) demonstrated the consistent quality of RPOF and showed a clear discrimination between RPOF and POFT. Principal component analysis, partial least-squares discriminant analysis, and heatmap-HCA on quantitative data not only gave a clear differentiation between RPOF and POFT, but they also suggested that quercetin, 3,5,7-trihydroxychromone, and N-trans-feruloyltyramine acted as the main factors responsible for the sample differences. Chromatographic analysis in combination with chemometric analysis provides a simple and reliable method of comparing and evaluating the qualities of RPOF and POFT.


The effect of cinnamon supplementation on glycemic control in women with polycystic ovary syndrome: A systematic review and meta-analysis.
Several clinical trials have identified glycemic-lowering effects of cinnamon, while other studies have reported conflicting findings. A comprehensive systematic search on Embase, PubMed, Scopus, Web of Science, and Cochrane Library was conducted using defined keywords in any language through June 2020. Studies that compared the effect of cinnamon with placebo on insulin resistance (IR) indices, as the primary outcome, in women with polycystic ovary syndrome (PCOS) were considered eligible. Standard Mean difference (SMD) (with 95% confidence intervals) for endpoints were calculated using the random-effects model. Finally, five RCTs which met the criteria were included in the meta-analysis. After pooling data, cinnamon supplementation significantly reduced homeostatic model assessment for insulin resistance (HOMA-IR) scores in women with PCOS (SMD: -0.84, 95% CI: -1.52, -0.16, p = .010). Cinnamon supplementation likely improves certain IR markers in patients with PCOS. PRACTICAL APPLICATIONS: There are controversies reports for cinnamon intake, which animal models have suggested that it decreases IR via promotion of insulin action, stimulating insulin signaling pathways, and enhancing insulin sensitivity. This study provides comprehensive information about the effect of cinnamon on insulin resistance (IR) indices in women with PCOS. In this regard, our results indicated that cinnamon supplementation significantly reduced homeostatic model assessment for insulin resistance (HOMA-IR) scores in women with PCOS. Therefore, consumption of cinnamon can be safe and this can be a useful recommendation for improving IR and promotion of healthy life which indeed are the potential or actual uses of this research.


Quasispecies analysis and in vitro susceptibility of HBV strains isolated from HIV-HBV-coinfected patients with delayed response to tenofovir.
Among 141 HIV-HBV-coinfected patients treated with tenofovir in our centre, 87% were good-responders to therapy. Seven patients showed a delayed response to tenofovir. The present study was performed to evaluate the quasispecies variability and the in vitro drug susceptibility to approved antiviral drugs of HBV genomes directly isolated from patients' sera. After purification of DNA from serum samples, full-length HBV DNA was amplified by PCR, cloned and sequenced. Drug sensitivity of HBV strains isolated from four delayed responders and five good-responders was assessed and compared to a wild-type HBV strain after transfection of the full genome into HepG2 cells. Delayed responders, compared with good responders, showed a higher incidence of lamivudine-resistant mutations (71% and 35%, respectively; P=0.021) and a higher proportion of HBV genotype G (57% versus 16%, respectively; P=0.026). Clonal analysis demonstrated a higher variability of HBV quasispecies in delayed reponders than in good responders. In vitro analysis showed a lower efficacy of adefovir and tenofovir in delayed reponders. Furthermore, HBV genotype G strains showed a mild to weak susceptibility to tenofovir. The reason for the slow decline in HBV DNA level observed during therapy in delayed responders is not clear. Delayed responders showed higher quasispecies variability, a higher proportion of HBV genotype G and a mild in vitro decreased susceptibility to tenofovir and adefovir. A combination of these factors in heavily treatment-experienced HIV-infected patients could explain the lower tenofovir activity. These patients must be closely monitored to prevent prospective emergence of resistance to approved antiviral drugs.


Mechanochemical degradation kinetics of high-density polyethylene melt and its mechanism in the presence of ultrasonic irradiation.
In this paper, the effect of ultrasonic intensity on the degradation of high-density polyethylene (HDPE) melt, degradation mechanism, ultrasonic degradation kinetics of HDPE melt as well as the development of molecular weight distribution of HDPE melt during ultrasonic degradation were studied. In the initial stage, the ultrasonic degradation of HDPE melt shows a random scission process, and the molecular weight distribution broadens. After that, the ultrasonic degradation of HDPE melt shows a nonrandom scission process, and the molecular weight distribution of HDPE melt narrows with ultrasonic irradiation time. The average molecular weight of HDPE decreases with the increase of ultrasonic intensity and increases and trends forward that of undegraded HDPE with the increase of distance from ultrasonic probe tip, indicating that attenuation of ultrasonic intensity in HDPE melt is very quick. Ultrasonic degradation kinetics of HDPE melt obeys the equation: Mt=M(infinity) + Ae(-kt). The theoretic calculation by this equation accords well with the experimental results. The plausible ultrasonic degradation mechanism of polymer melt based on molecular relaxation was also proposed in this paper.


The relationship between anabolic androgenic steroids and muscle dysmorphia: a review.
This review explores the condition of muscle dysmorphia (MD) and its relationship with anabolic androgenic steroids (AAS). Particular emphasis is placed upon whether anabolic steroids are a predisposing, precipitating or perpetuating factor of MD. Furthermore, psychiatric complications of AAS abuse are examined. The current evidence from the literature suggests that AAS (ab)use is possibly a perpetuating factor in the evolution of MD. Psychiatric complications of AAS include mood and behavior changes, perceptual abnormalities, and withdrawal symptoms. In addition, there appears to be a credible dependence theory to AAS in fruition.


Photolytic transformation products and biological stability of the hydrological tracer Uranine.
Among many fluorescence tracers, Uranine (sodium fluorescein, UR) has most widely been used in hydrological research. Extensive use of UR for tracing experiments or commercial use might cause a potential risk of long-term environmental contamination. As any organic substance released to the environment, also UR is subjected to chemical and physical reactions that can be chemical, biological and photolysis processes. These processes transform the parent compound (PC) and have not been extensively investigated for UR. This study applies two OECDs (301 D and 301 F) tests and a screening water sediment test (WST) to investigate the biodegradability of the PC. Photolysis in water was explored by Xe lamp irradiation. Subsequently, the biodegradability of the photolysis mixtures was examined. The primary elimination of UR was monitored and structures of its transformation products (TPs) were elucidated by HPLC-FLD-MS/MS. UR was found not readily biodegradable, although small degradation rates could be observed in the OECD 301 D and WST. HPLC-FLD analysis showed high primary elimination of the tracer during photolysis. However, the low degree of mineralization found indicates that the UR was not fully degraded, instead transformed to TPs. A total of 5 photo-TPs were identified. According to MS/MS data, chemical structures could be proposed for all identified photo-TPs. Likewise the parent compound it was demonstrated that photo-TPs were largely recalcitrant to microbial degradation. Although we did not find indications for toxicity, target-oriented studies on the environmental impact of these photo-TPs are warranted. Results obtained in this study show that deeper investigations are necessary to fully understand fate and risk connected to the use of UR.


[Genetics of neurodevelopmental disorders. Practical aspects].
Neurodevelopmental disorders (NDD) constitute a relevant group of pathologies, of childhood, with a biological basis and totally or partially genetic etiology. The recognition of the causal factors constitutes a challenge that has been perfected over the last decades, until obtaining an increasing diagnostic yield. The implementation of these technological advances can only be achieved through the formation of interdisciplinary work teams, which, following an orderly process, achieve a presumptive diagnosis, which is then certified using the techniques that for each of the cases are more profitable in terms of quality and cost. In this paper we list these procedures, based on different scenarios that highlight the extensive menu of possibilities and the need to manage them in a rational way, on well-founded scientific bases.


Electrophysiological characterization of responses from gustatory receptor neurons of sensilla chaetica in the moth Heliothis virescens.
Discrimination of edible and noxious food is crucial for survival in all organisms. We have studied the physiology of the gustatory receptor neurons (GRNs) in contact chemosensilla (insect gustatory organs) located on the antennae of the moth Heliothis virescens, emphasizing putative phagostimulants and deterrents. Sucrose and the 2 bitter substances quinine and sinigrin elicited responses in a larger proportion of GRNs than inositol, KCl, NaCl, and ethanol, and the firing thresholds were lowest for sucrose and quinine. Variations in GRN composition in individual sensilla occurred without any specific patterns to indicate specific sensillum types. Separate neurons showed excitatory responses to sucrose and the 2 bitter substances quinine and sinigrin, implying that the moth might be able to discriminate bitter substances in addition to separating phagostimulants and deterrents. Besides being detected by separate receptors on the moth antennae, the bitter tastants were shown to have an inhibitory effect on phagostimulatory GRNs. Sucrose was highly appetitive in behavioral studies of proboscis extension, whereas quinine had a nonappetitive effect in the moths.


Neuromuscular electrical stimulation reduces skeletal muscle protein degradation and stimulates insulin-like growth factors in an age- and current-dependent manner: a randomized, controlled clinical trial in major abdominal surgical patients.
To investigate the effect of neuromuscular electrical stimulation (NMES) on skeletal muscle metabolism after major abdominal surgery. Protein catabolism associated with surgical interventions leads to reduced muscle strength, increased clinical complications and prolonged convalescence. Immobilization is suggested as a major stimulus for muscle wasting. This study investigates the potency of NMES on skeletal muscle growth factors and degradation processes in surgical patients. This observer blind study included 26 patients after major abdominal surgery mainly due to cancer aged 60 +/- 10 years. Starting on the first postoperative day, 1 randomly assigned thigh of each patient was treated on 4 consecutive days with NMES, whereas the other leg was used as sham-stimulated control. Thereafter, muscle biopsies from both legs were performed. Differences in mRNA level, protein expression, and enzyme activity between legs were analyzed by cross-over analysis of variance (Clinical Trial Registration Number: NCT00635440). NMES significantly increased total RNA content and total sarcoplasmatic protein content. NMES significantly reduced ubiquitin-conjugated sarcoplasmatic proteins and proteasome activity. The mechano growth factor mRNA level correlated positively with the applied current and negatively with the body mass index of the patients. The increase in insulin like growth factor-1Ea mRNA after NMES correlated negatively with the age of the patients. This study shows that NMES significantly increases total RNA content and reduces protein degradation in postoperative patients. Moreover, the induction of growth factors by NMES reveals dependency on body mass index, age, and applied current. We conclude that NMES is a useful clinical tool to reduce protein catabolism in postoperative patients.


Afghanistan: demographic consequences of war, 1978-1987.
"In this paper, I attempt to analyse the effects of war on the size and age-sex structure of the Afghan population, and to estimate the size of the Afghan population in Afghanistan by age and sex for the years 1978-1987. In doing so, I try to estimate the number of persons killed during the war and examine the validity of the available estimates." The analysis is based on data from the 1979 census, a 1987 survey, and other sources.


Role of mitogen-activated protein kinase in osteoblast differentiation.
Local control of osteoblast differentiation and bone formation is not well understood. We have previously seen biphasic effects on cell differentiation in response to the short- and long-term exposure to IL-1β in rat calvarial osteoblasts. To characterize the signaling pathway mechanisms regulating IL-1β biphasic effects, we examined the contribution of mitogen-activated protein kinase (MAPK) family. Cells were pretreated with specific inhibitors to extracellular signal-regulated kinase (ERK, PD98059), p38 (SB203580), and c-JUN N-terminal kinase (JNK, SP600125), then co-cultured with IL-1β for 2, 4, and 6 days. Cell differentiation was determined by measuring bone nodules after 10 days of culture. These inhibitors did not alter biphasic effects of IL-1β on cell differentiation. However, PD98059 and U2016, another inhibitor of ERK activation robustly increased osteoblast differentiation compared to vehicle-treated control in a time- and dose-dependent manner. PD98059 appears to stimulate alkaline phosphatase (ALP) activity to promote cell differentiation, where IL-1β appears to suppress it. Interestingly, continuous ERK inhibition with PD98059, after 2 and 4 days of IL-1β treatment, enhanced the IL-1β anabolic effect by increasing bone nodules formed. These observations provide a potential mechanism involving ERK pathway in osteoblasts differentiation and suggest that MAPK family may not directly regulate IL-1β biphasic effects.


Femoral Positioning of the Anterolateral Ligament Graft With and Without Ultrasound Location of the Lateral Epicondyle.
In anterior cruciate ligament (ACL) reconstruction with anterolateral ligament (ALL) reconstruction, precise positioning of the ALL graft on the femur and tibia is key to achieve rotational control. The lateral femoral epicondyle is often used as a reference point for positioning of the ALL graft and can be located by palpation or with ultrasound guidance. To compare the ALL graft positioning on the femoral side between an ultrasound-guided technique and a palpation technique for the location of the lateral epicondyle. Cohort study; Level of evidence, 2. A total of 120 patients receiving a primary combined ACL and ALL reconstruction between June and December 2019 were included. The location of the lateral epicondyle was determined by palpation in the palpation group (n = 60) and with preoperative ultrasound guidance in the ultrasound group (n = 60). Groups were comparable in age, sex, body mass index (BMI), and operated side. The planned positioning of the femoral ALL graft was proximal and posterior to the lateral epicondyle. The effective positioning of the femoral ALL graft was evaluated on postoperative lateral radiographs. The primary outcome was location of the graft in a 10-mm quadrant posterior and proximal to the lateral epicondyle. Results were analyzed in 2 subgroups according to BMI. All 60 anterolateral grafts (100%) in the ultrasound group were positioned in a 10-mm quadrant posterior and proximal to the lateral epicondyle, as opposed to 52 (87%) in the palpation group (P = .006). Errors in graft positioning with palpation occurred in overweight patients (BMI >25) as well as nonoverweight patients (P = .3). Femoral positioning of the ALL graft posterior and proximal to the lateral epicondyle is more reproducible with ultrasound guidance when compared with palpation alone, regardless of BMI.


Wound botulism--California, 1995.
During January-November 1995, a total of 19 laboratory-confirmed cases of wound botulism were reported to the California Department of Health Services (CDHS); of these, 13 had occurred since August. Since 1990, the number of wound botulism cases reported annually in California has increased steadily (one case in 1990, two in 1991, three in 1992, four in 1993, and 11 in 1994). All cases except one since 1991 have occurred in injecting-drug users, and many involved subcutaneous injection or "skin popping" of black tar heroin. This report summarizes the findings of the investigation of two cases.


Tripping the reset switch.
Simon Toye and his wife, who is a vet, are just back from a trip through Asia. He says it was a journey that bolstered their wellbeing and left them better equipped to deal with the pressures of veterinary practice.


Left ventricular dysfunction during dobutamine stress echocardiography in patients with syndrome X and positive myocardial perfusion scintigraphy.
A sizeable proportion of patients with angina, angiographically smooth coronary arteries and positive exercise test (syndrome X) have stress/rest myocardial perfusion defects. The aim of the study was to assess whether perfusion defects are dependent upon a reduction in coronary flow reserve causing regional left ventricular dysfunction in syndrome X patients. Twenty-two syndrome X patients underwent dobutamine stress echocardiography (DSE). All had stress-induced perfusion defects documented by 99m-Tc-MIBI scintigraphy. Resting and peak DSE wall motion score index (WMSI) were evaluated. Six patients exhibited resting wall motion abnormalities in 10 segments (WMSI 1.05 +/- 0.11). DSE was positive in 12 patients (53%), in whom 16 myocardial segments were involved: of these, 12 were normokinetic and 4 hypokinetic at rest. Peak WMSI was 1.17 +/- 0.17 (p < 0.05 vs rest). Of the 12 patients with a positive DSE, 9 also showed diagnostic ECG changes and 6 complained of angina. Of the 10 patients with negative DSE, 5 had angina and 5 (one with angina) showed ECG changes. In 7 patients (7 segments) (32%), the location of dobutamine-induced wall motion abnormalities coincided with the area where exercise-induced hypoperfusion was observed with MIBI. More than a half of syndrome X patients with myocardial perfusion abnormalities also develop regional LV dysfunction during DSE. However, the site of perfusion defects and wall motion abnormalities can be different. Reversible ischemia, defined as a parallel limitation of flow reserve and inducible dysfunction, could be identified as the cause of chest pain in almost one-third of patients.


Galleria mellonella as an infection model for the virulent Mycobacterium tuberculosis H37Rv.
Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), is a leading cause of infectious disease mortality. Animal infection models have contributed substantially to our understanding of TB, yet their biological and non-biological limitations are a research bottleneck. There is a need for more ethically acceptable, economical, and reproducible TB infection models capable of mimicking key aspects of disease. Here, we demonstrate and present a basic description of how Galleria mellonella (the greater wax moth, Gm) larvae can be used as a low cost, rapid, and ethically more acceptable model for TB research. This is the first study to infect Gm with the fully virulent MTB H37Rv, the most widely used strain in research. Infection of Gm with MTB resulted in a symptomatic lethal infection, the virulence of which differed from both attenuated Mycobacterium bovis BCG and auxotrophic MTB strains. The Gm-MTB model can also be used for anti-TB drug screening, although CFU enumeration from Gm is necessary for confirmation of mycobacterial load reducing activity of the tested compound. Furthermore, comparative virulence of MTB isogenic mutants can be determined in Gm. However, comparison of mutant phenotypes in Gm against conventional models must consider the limitations of innate immunity. Our findings indicate that Gm will be a practical, valuable, and advantageous additional model to be used alongside existing models to advance tuberculosis research.


Regression-based noninvasive estimation of intracranial pressure.
Monitoring of intracranial pressure (ICP) is indicated in patients with a variety of conditions affecting the brain and cerebrospinal fluid space. The measurement of ICP, however, is highly invasive as it requires placement of a catheter in the brain tissue or cerebral ventricular spaces. Several noninvasive techniques have been proposed to overcome this issue, and one class of approaches is based on analyzing cerebral blood flow velocity (CBFV) and arterial blood pressure (ABP) waveforms to infer ICP. Here, we analyze a physiologic model linking ICP to CBFV and ABP and present a regression-based approach to estimating ICP. We tested the model on 20 datasets recorded from three patients in intensive care. Our estimates achieve a mean error (bias) of -1.12 mmHg and a standard deviation of the error of 5.56 mmHg, for a root-mean-square error of 5.68 mmHg, when compared against the invasive ICP measurement. Since transcranial Doppler ultrasound based CBFV measurements depend on the Doppler angle φ between the direction of the ultrasound beam and the (main) direction of blood flow velocity, we investigated the robustness of our ICP estimates against variations in φ. Our results show a change in the estimated ICP that is <;1 mmHg if we assume φ ~ N(μ; σ2), with μ = 0 and σ = 10°.