Structure-based thermodynamics of ion selectivity (Mg2+versus Ca2+ and K+versus Na+) in the active site of the eukaryotic lariat group II intron from algae Pylaiella littoralis.
Group II introns are metalloenzymes that can catalyze self-splicing. Recently, the crystal structures of the eukaryotic group IIB lariat intron from the brown algae Pylaiella littoralis have been reported for two intermediate states (pre-hydrolytic (2s) and post-hydrolytic) along the self-splicing pathway. Three characteristic metal-ion binding sites (M1 and M2 sites for catalytic Mg2+ ions, and K1 site for K+) in the catalytic pocket of the lariat intron have been identified and proposed to be crucial for self-splicing. Using the X-ray structures as a template, we quantitatively estimated the energetics of divalent (Mg2+versus Ca2+) and monovalent (K+versus Na+) ion selectivity and established a direct link between the energetics and structures of this lariat intron (bound to cognate and near-cognate metal ions). Molecular dynamics (MD) free energy simulations showed that the lariat intron was strongly selective between divalent metal ions. The strength of divalent metal-ion selectivity was noticeably high in the post-hydrolytic state (ΔΔG ≈ 20 kcal mol-1) relative to its pre-hydrolytic (2s) state (ΔΔG ≈ 13 kcal mol-1). Quantum chemical calculations ensured that the sign of the estimated divalent metal-ion selectivity was correct. The M1-binding pocket was less solvent-exposed in the case of the post-hydrolytic state relative to the pre-hydrolytic (2s) state, which boosted the metal-ion selectivity of the former. Surprisingly, in contrast to the bacterial linear group II intron, the lariat intron was found to be non-selective between monovalent ions (K+versus Na+). The interaction network in the first coordination shell of Ca2+ in the M1-binding pocket was different relative to Mg2+. Mg2+ substitution by Ca2+ resulted in the substitution of a single M1-RNA interaction by the M1-water interaction. In the pre-hydrolytic (2s) state, Ca2+ substitution completely disrupted the M1⋯5'-exon interaction; thus, the nature of the divalent metal ion is critical for catalysis. The interaction network in the M2 site was independent of the nature of the divalent metal ions (Mg2+ or Ca2+). The monovalent ion was loosely bound in the wet binding pocket (K1 site) of the lariat intron; thus, the substitution of K+ by Na+ could not significantly alter the free energy of the complex. The metal ion selectivity was dependent on the solvent accessibility of the metal-ion-binding-pocket, dry pocket enhanced the selectivity.


Variations in Knee Kinematics After ACL Injury and After Reconstruction Are Correlated With Bone Shape Differences.
The factors that contribute to the abnormal knee kinematics after anterior cruciate ligament (ACL) injury and ACL reconstruction remain unclear. Bone shape has been implicated in the development of hip and knee osteoarthritis, although there is little knowledge about the effects of bone shape on knee kinematics after ACL injury and after ACL reconstruction. QUESTIONS/QUESTIONS: (1) What is the relationship between bony morphology with alterations in knee kinematics after ACL injury? (2) Are baseline bone shape features related to abnormal knee kinematics at 12 months after ACL reconstruction? Thirty-eight patients (29 ± 8 years, 21 men) were prospectively followed after acute ACL injury and before ligamentous reconstruction. Patients were excluded if there was a history of prior knee ligamentous injury, a history of inflammatory arthritis, associated meniscal tears that would require repair, or any prior knee surgery on either the injured or contralateral side. In total, 54 patients were recruited with 42 (78%) patients completing 1-year followup and four patients excluded as a result of incomplete or unusable imaging data. MR images were obtained for the bilateral knees at two time points 1 year apart for both the injured (after injury but before reconstruction and 1 year after reconstruction) and contralateral uninjured knees. Kinematic MRI was performed with the knee loaded with 25% of total body weight, and static images were obtained in full extension and in 30° of flexion. The side-to-side difference (SSD) between tibial position in the extended and flexed positions was determined for each patient. Twenty shape features, referred to as modes, for the tibia and femur each were extracted independently from presurgery scans with the principal component analysis-based statistical shape modeling algorithm. Spearman rank correlations were used to evaluate the relationship between the SSD in tibial position and bone shape features with significance defined as p < 0.05. Each of the shape features (referred to as the bone and mode number such as Femur 18 for the 18th unique femoral bone shape) associated with differences in tibial position was then investigated by modeling the mean shape ± 3 SDs. Two of the 20 specific femur bone shape features (Femur 10, Femur 18) and two of the 20 specific tibial bone shape features (Tibia 19, Tibia 20) were associated with an increasingly anterior SSD in the tibial position for the patients with ACL injury before surgical treatment. The shape features described by these modes include the superoinferior height of the medial femoral condyle (Femur 18; ρ = 0.33, p = 0.040); the length of the anterior aspect of the lateral tibial plateau (Tibia 20; ρ = -0.35, p = 0.034); the sphericity of the medial femoral condyle (Femur 10; ρ = -0.52, p < 0.001); and tibial slope (Tibia 19; ρ = 0.34; p = 0.036). One year after surgical treatment, there were two of 20 femoral shape features that were associated with SSD in the tibial position in extension (Femur 10, Femur 18), one of 20 femoral shape features associated with SSD in the tibial position in flexion (Femur 10), and three of 20 tibial shape features associated with SSD in the tibial position in flexion (Tibia 2, Tibia 4, Tibia 19). The shape features described by these modes include the sphericity of the medial femoral condyle (Femur 10; ρ = -0.38, p = 0.020); the superoinferior height of the medial femoral condyle (Femur 18; ρ = 0.34, p = 0.035); the height of the medial tibial plateau (Tibia 2; ρ = -0.32, p = 0.048); the AP length of the lateral tibial plateau (Tibia 4; ρ = -0.37, p = 0.021); and tibial slope (Tibia 19; ρ = 0.34, p = 0.038). We have observed multiple bone shape features in the tibia and the femur that may be associated with abnormal knee kinematics after ACL injury and ACL reconstruction. Future directions of research will include the influence of bony morphology on clinical symptoms of instability in patients with and without ACL reconstruction and the long-term evaluation of these shape factors to better determine specific contributions to posttraumatic arthritis and graft failure. Level II, therapeutic study.


Retinoic acid conjugates as potential antitumor agents: synthesis and biological activity of conjugates with Ara-A, Ara-C, 3(2H)-furanone, and aniline mustard moieties.
In a dual targeting approach, to explore the ability of tretinoin (all-trans-retinoic acid) to behave as a covalent carrier for cytotoxic entities, conjugates of retinoic acid with a few representative molecules, being important examples of antitumor pharmacophores (i.e., nucleoside analogues and alkylating agents), have been synthesized and tested for their cytostatic and differentiating activity. All compounds were stable to in vitro hydrolysis in human plasma and more lipophilic than the parent compounds, thus consenting enhanced uptake into the cells. Among the nucleoside analogues the Ara-C derivatives 3 and 6 and the Ara-A derivative 7 proved the most cytostatic (IC50 < 0.32 microgram/mL) resulting from 25- to > 144-fold more active (Ara-A derivatives) or at least as equally active (Ara-C derivatives) as compared to the parent nucleosides. Compound 3, endowed with a highly lipophilic silyl moiety at the 3' and 5' positions, showed the highest differentiating activity (54% and 44% differentiated HL-60 cells at 0.2 and 0.05 microgram/mL respectively). With regard to the retinoic acid conjugates of alkylating agents, compound 10 was the most cytostatic agent (IC50 < 0.32 microgram/mL) and the most potent differentiating agent (33-34% at 0.32 and 0.08 microgram/mL). These structures may also be regarded as analogs of either retinoic acid or the cytotoxic compound.


Combined Economic Emission Dispatch of Microgrid with the Incorporation of Renewable Energy Sources Using Improved Mayfly Optimization Algorithm.
Electricity can be provided to small-scale communities like commercial areas and villages through microgrid, one of the small-scale, advanced, and independent electricity systems out of the grid. Microgrid is an appropriate choice for specific purposes reducing emission and generation cost and increasing efficiency, reliability, and the utilization of renewable energy sources. The main objective of this paper is to elucidate the combined economic emission dispatch CEED problem in the microgrid to attain optimal generation cost. A combined cost optimization approach is examined to minimize operational cost and emission levels while satisfying the load demand of the microgrid. With this background, the authors proposed a novel improved mayfly algorithm incorporating Levy flight to resolve the combined economic emission dispatch problem encountered in microgrids. The islanded mode microgrid test system considered in this study comprises thermal power, solar-powered, and wind power generating units. The simulation results were considered for 24 hours with varying power demands. The minimization of total cost and emission is attained for four different scenarios. Optimization results obtained for all scenarios using IMA give a comparatively better reduction in system cost than MA and other optimization algorithms considered revealing the efficacy of IMA taken for comparison with the same data. The proposed IMA algorithm can solve the CEED problem in a grid-connected microgrid.


Protein kinase C alpha/beta inhibitor Go6976 promotes formation of cell junctions and inhibits invasion of urinary bladder carcinoma cells.
Changes in activation balance of different protein kinase C (PKC) isoenzymes have been linked to cancer development. The current study investigated the effect of different PKC inhibitors on cellular contacts in cultured high-grade urinary bladder carcinoma cells (5637 and T24). Exposure of the cells to isoenzyme-specific PKC inhibitors yielded variable results: Go6976, an inhibitor of PKCalpha and PKCbeta isoenzymes, induced rapid clustering of cultured carcinoma cells and formation of an increased number of desmosomes and adherens junctions. Safingol, a PKCalpha inhibitor, had similar but less pronounced effects. In contrast, a PKCdelta inhibitor, rottlerin, had an opposite effect on cell clustering and caused dissociation of cell junctions. A broad-spectrum PKC inhibitor bisindolylmaleimide I did not have any apparent effect on the morphology of the cultures or on the number of cell junctions. Additional studies with Go6976 demonstrated that inhibition of PKCalpha and beta isoenzymes induced translocation of beta1-integrin from the cell-matrix junctions and that beta4-integrin was translocated to face the culture substratum. Go6976 was also highly effective in inhibiting migration of carcinoma cells and inhibited invasion through artificial basement membrane. Our results on urinary bladder carcinoma cells emphasize that Go6976 is a potential anticancer drug due to its effects on cell-cell and cell-matrix junctions, migration, and invasion. Furthermore, the results may be explained by changes in PKC activation balance promoted by inhibition of PKCalpha/beta.


Somatostatin Ameliorates β-Amyloid-Induced Cytotoxicity via the Regulation of CRMP2 Phosphorylation and Calcium Homeostasis in SH-SY5Y Cells.
Somatostatin is involved in the regulation of multiple signaling pathways and affords neuroprotection in response to neurotoxins. In the present study, we investigated the role of Somatostatin-14 (SST) in cell viability and the regulation of phosphorylation of Collapsin Response Mediator Protein 2 (CRMP2) (Ser522) via the blockade of Ca2+ accumulation, along with the inhibition of cyclin-dependent kinase 5 (CDK5) and Calpain activation in differentiated SH-SY5Y cells. Cell Viability and Caspase 3/7 assays suggest that the presence of SST ameliorates mitochondrial stability and cell survival pathways while augmenting pro-apoptotic pathways activated by Aβ. SST inhibits the phosphorylation of CRMP2 at Ser522 site, which is primarily activated by CDK5. Furthermore, SST effectively regulates Ca2+ influx in the presence of Aβ, directly affecting the activity of calpain in differentiated SH-SY5Y cells. We also demonstrated that SSTR2 mediates the protective effects of SST. In conclusion, our results highlight the regulatory role of SST in intracellular Ca2+ homeostasis. The neuroprotective role of SST via axonal regeneration and synaptic integrity is corroborated by regulating changes in CRMP2; however, SST-mediated changes in the blockade of Ca2+ influx, calpain expression, and toxicity did not correlate with CDK5 expression and p35/25 accumulation. To summarize, our findings suggest two independent mechanisms by which SST mediates neuroprotection and confirms the therapeutic implications of SST in AD as well as in other neurodegenerative diseases where the effective regulation of calcium homeostasis is required for a better prognosis.


Biological targets of antitumor indolocarbazoles bearing a sugar moiety.
Natural and synthetic indolocarbazole compounds have triggered considerable interest since the discovery in 1986 of the inhibitory properties of staurosporine toward protein kinase C (PKC). Later, it has been shown that indolocarbazole compounds may inhibit various kinases, such as cyclin dependent-kinases and/or topoisomerase I, someones behave only as DNA intercalators. In this review are presented various indolocarbazole compounds bearing a sugar moiety and their biological targets. The relevance of these targets to develop indolocarbazole compounds as potential antitumor agents is discussed.


Photodynamic therapy for central serous chorioretinopathy.
Central serous chorioretinopathy (CSCR) is an idiopathic disorder characterised by detachment of the neurosensory retina due to serous fluid accumulation between the photoreceptor outer segments and the retinal pigment epithelium. There are currently no set guidelines or protocols on its treatment. This study was undertaken to assess the current literature on the the efficacy and safety of photodynamic therapy (PDT) as a treatment option for CSCR. Seven databases (PubMed, CENTRAL, MEDLINE, Web of Science, Embase, Scopus, and The Cochrane Database of Systematic Reviews) were searched without restrictions on time or location. We followed PRISMA guidelines and evaluated quality according to STROBE criteria. In total, 117 citations were identified and 31 studies describing 787 eyes were included for review. Data on indications for PDT in CSCR, dosing regimens of verteprofin PDT (which includes treatment dose of vertoporfin, treatment time, fluence, and spot size), number of treatment sessions, response to treatment, mean length of follow-up, and complications were extracted and analysed. Since the introduction of PDT for the treatment of CSCR in 2003, there have been three randomised controlled trials (RCTs), one for acute and two chronic CSCR and 28 further studies that met the STROBE criteria that compared the use of PDT with other treatment options. All studies showed short-term efficacy of PDT in CSCR. The studies were of small sample size and lacked sufficient follow-up to draw conclusions on long-term efficacy and safety. There is sufficient scientific evidence to suggest that PDT may be a useful treatment option for chronic CSCR in the short-term. The review identifies a need for robust RCTs with longer follow-up to ascertain the role of PDT as a useful treatment option for CSCR.


[The clinical pharmacology of antiepileptic agents].
The correct choice of an antiepileptic drug, its individual dosage and schedule of administration at the beginning of treatment and then during the maintenance therapy promotes a complete control of seizures using one anticonvulsant in 60-90% of patients. The treatment should be initiated with administration of one drug which is adequate for the given type of seizures taking into consideration that different drugs can appear effective in different patients with the same form of epilepsy. One has to choose drugs by consecutively gradually changing them. The optimization of epilepsy treatment should be performed on the basis of the knowledge of the clinicopharmacological characteristic of the main antiepileptic drugs described in the lecture.


Cognitive-behavioral treatment for chronic nightmares in trauma-exposed persons: assessing physiological reactions to nightmare-related fear.
Cognitive-behavioral treatments (CBTs) that target nightmares are efficacious for ameliorating self-reported sleep problems and psychological distress. However, it is important to determine whether these treatments influence objective markers of nightmare-related fear, because fear and concomitant physiological responses could promote nightmare chronicity and sleep disturbance. This randomized, controlled study (N=40) assessed physiological (skin conductance, heart rate, facial electromyogram) and subjective (displeasure, fear, anger, sadness, arousal) reactions to personally relevant nightmare imagery intended to evoke nightmare-related fear. Physiological assessments were conducted at pretreatment as well as 1-week, 3-months, and 6-months posttreatment. Results of mixed effects analysis of variance models suggested treatment reduced physiological and subjective reactions to nightmare imagery, gains that were generally maintained at the 6-month follow-up. Potential implications are discussed.


Neuroprotective Effects of a Hydrogen Sulfide Donor in Streptozotocin-Induced Diabetic Rats.
Diabetic neuropathy is an important long-term complication of diabetes. This study explored the hypothesis that hydrogen sulfide (H2S) ameliorates neuropathic pain by controlling antiapoptotic and pro-apoptotic processes. The effects of a slow-releasing H2S donor, GYY4137, on the expression of antiapoptotic and pro-apoptotic genes and proteins, such as B-cell lymphoma 2 (Bcl2) and Bcl-2-like protein 4 (Bax), as well as caspases, cyclooxygenase (COX)-1 and COX-2, monocytes/macrophages, and endothelial cells, in the spinal cord of male Sprague-Dawley rats with streptozotocin-induced peripheral diabetic neuropathy, were investigated using reverse transcription-PCR, western blot and immunohistochemistry. The antihypoalgesic activities of GYY4137 on diabetic rats were evaluated using the tail flick test. Treatment of diabetic rats with GYY4137 attenuated thermal hypoalgesia and prevented both the diabetes-induced increase in Bax mRNA expression (p = 0.0032) and the diabetes-induced decrease in Bcl2 mRNA expression (p = 0.028). The GYY4137-treated diabetic group had increased COX-1 (p = 0.015), decreased COX-2 (p = 0.002), reduced caspase-7 and caspase-9 protein expression (p < 0.05), and lower numbers of endothelial and monocyte/macrophage cells (p < 0.05) compared to the non-treated diabetic group. In summary, the current study demonstrated the protective properties of H2S, which prevented the development of neuropathy related behavior, and suppressed apoptosis activation pathways and inflammation in the spinal cord. H2S-releasing drugs could be considered as possible treatment options of diabetic peripheral neuropathy.


Development of a Questionnaire and Cross-Sectional Survey of Patient eHealth Readiness and eHealth Inequalities.
Many speak of the digital divide, but variation in the opportunity of patients to use the Internet for health (patient eHealth readiness) is not a binary difference, rather a distribution influenced by personal capability, provision of services, support, and cost. Digital divisions in health have been addressed by various initiatives, but there was no comprehensive validated measure to know if they are effective that could be used in randomized controlled trials (RCTs) covering both non-Internet-users and the range of Internet-users. The aim of this study was to develop and validate a self-completed questionnaire and scoring system to assess patient eHealth readiness by examining the spread of scores and eHealth inequalities. The intended use of this questionnaire and scores is in RCTs of interventions aiming to improve patient eHealth readiness and reduce eHealth inequalities. Based on four factors identified from the literature, a self-completed questionnaire, using a pragmatic combination of factual and attitude questions, was drafted and piloted in three stages. This was followed by a final population-based, cross-sectional household survey of 344 people used to refine the scoring system. the patient ehealth readiness questionnaire (perq) includes questions used to calculate four subscores: patients' perception of (1) provision, (2) their personal ability and confidence, (3) their interpersonal support, and (4) relative costs in using the Internet for health. These were combined into an overall PERQ score (0-9) which could be used in intervention studies. Reduction in standard deviation of the scores represents reduction in eHealth inequalities. PERQ appears acceptable for participants in British studies. The scores produced appear valid and will enable assessment of the effectiveness of interventions to improve patient eHealth readiness and reduce eHealth inequalities. Such methods need continued evolution and redevelopment for other environments. Full documentation and data have been published to allow others to develop the tool further.


Calcium antagonists prevent monocyte and endothelial cell-induced modification of low density lipoproteins.
Low density lipoprotein (LDL) incubated in the presence of the calcium antagonists verapamil, nifedipine and flunarizine were more resistant than control LDL to human monocyte- or endothelial cell-induced modification, as assessed by electrophoretic mobility in agarose gel, thiobarbituric acid reactive substance content, and degradation by J774 macrophages. The efficiency of the drugs was: flunarizine greater than nifedipine greater than verapamil. Moreover, a 24 h preculture with calcium antagonists significantly impaired the ability of cells to modify LDL in the absence of the drugs. All the studied drugs also inhibited copper-induced autooxidation of LDL. None of the studied calcium antagonists, at concentrations up to 10(-4) M, significantly reacted with free radicals as assessed by the 1,1-diphenyl-2-picrylhydrazyl test. It is suggested that such a protective effect of calcium antagonists against LDL peroxidation could play a role in the previously reported antiatherogenic effect of these drugs.


Synthesis, spectral characterization, and in vitro cellular activities of metapristone, a potential cancer metastatic chemopreventive agent derived from mifepristone (RU486).
Mifepristone (RU486) is marketed and used widely by women as an abortifacient, and experimentally for psychotic depression and anticancer treatments. After administration, metapristone is found to be the most predominant metabolite of mifepristone. We hypothesized that adhesion of circulating tumor cells (CTCs) to vascular endothelial bed is a crucial starting point in metastatic cascade, and that metapristone can serve as a cancer metastatic chemopreventive agent that can interrupt adhesion and invasion of CTCs to the intima of microvasculature. In the present study, we modified the synthesis procedure to produce grams of metapristone, fully characterized its spectral properties and in vitro cellular activities, including its cytostatic effects, cell cycle arrest, mitochondrial membrane potential, and apoptosis on human colorectal cancer HT-29 cells. Metapristone concentration dependently interrupted adhesion of HT-29 cells to endothelial cells. Metapristone may potentially be a useful agent to interrupt metastatic initiation.


The value of cancer antigen-125 as a tumor marker in malignant germ cell tumors of the ovary.
The value of cancer antigen-125 (CA-125) as a tumor marker for malignant germ cell tumors (MGCT) of the ovary was investigated and compared with the other recognized tumor markers (human chorionic gonadotrophin (hCG), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA) and lactate dehydrogenase (LDH) isoenzymes. In the 10 months following June 1984, 4 new cases with MGCT and 1 patient with active disease on treatment were evaluated. In all cases prior to planned surgery the levels of CA-125 were significantly elevated. The serum values ranged from 154 to 617 U/ml (normal less than 20 U/ml). In 1 case (pure dysgerminoma) CA-125 was the only tumor marker. In 3 patients (2 mixed germ cell tumors and 1 immature teratoma) serum LDH (LD 1, 2, and 3) was elevated, and AFP was elevated in 1 of these. In the fifth case (mixed germ cell tumor), on treatment, serum AFP was used to monitor the disease. Four patients underwent cytoreductive surgery followed by combination chemotherapy. The changes in the serum levels of CA-125 paralleled those of the other tumor markers while on therapy. In our experience CA-125 is an invaluable indicator of the clinical status of the patient and could be a new tumor marker in patients with MGCT.


[Swimming as a means of enhancing the adaptive potentials of the disabled after amputation of the lower extremities].
The studies of adaptive-compensatory reactions in low-limb amputees on training swimming regimens registered changes in vegetative regulation, predominance of the sympathetic component, low cardiovascular adaptive potential. High amputations led to arterial hypertension. Swimming and underwater exercise suppressed the influence of the sympathetic component of the autonomic nervous system, promoted enhancement of automatic regulation of the cardiac rhythm, lowering of arterial pressure. Pool exercise are recommended for rehabilitation of amputees.


[Emergency endovascular treatment of acute aortic diseases].
To investigate the safety and efficacy of emergency treatment of acute aortic diseases with endovascular stent grafts. In 11 patients (median age 55 years, range 18 - 85) with acute complications of descending aortic diseases endovascular emergency treatment was performed: traumatic aortic rupture (n = 4), penetrating ulcer with aortobronchial fistula or hematothorax (n = 4), acute type B dissection (n = 2, one with penetration, one with subacute mesenteric ischemia), and symptomatic aneurysm of the thoracic aorta (n = 1) with pain and diameter progression. 15 stent grafts were implanted (Talent n = 11, Vanguard/Stentor n = 4). Stent extension was necessary in 4 cases. In 2 cases graft extension was done during the first procedure (due to distal migration and due to the total length of the aortic aneurysm). In 2 cases graft extension was performed 5 days (due to a new aortic ulcer at the proximal stent struts) and 5 months after the initial procedure (recurrent aortobronchial fistula due to aneurysm progression). 14 of 15 implantations required general anesthesia, one symptomatic thoracic aneurysm was performed in local anesthesia and sedation. 14 of 15 graft procedures were performed using the femoral or iliac approach. One procedure required aortofemoral bypass grafting due to extensive arteriosclerotic stenosis and the stent graft was inserted via the bypass graft. The orifice of the subclavian artery was crossed with bare stent struts in 4 cases without neurological complications. Median follow-up is 27 months (range 6 to 72 months). In traumatic aortic ruptures, immediate sealing of bleeding was achieved and follow-up is inconspicuous at a maximum of 72 months. In cases of aortobronchial fistulas, follow-up is satisfactory (maximum 72 months) despite the necessity for reintervention and graft extension. In one acute type B dissection retrograde dissection of the aortic arch occurred during stent release with stable disease during follow-up without neurological complications. In one type B dissection with mesenteric ischemia the mesenteric blood flow was restored. A second look operation confirmed pulsatile flow in the mesenteric trunk but a total necrosis of the small intestine and the patient consequently died. Endovascular treatment is safe and effective for emergency treatment of life-threatening complications in selected acute aortic syndromes. Mid-term results are encouraging, however, regular follow-up is mandatory to recognize late complications of the stent graft.


Inhibitions of HMGB1 and TLR4 alleviate DINP-induced asthma in mice.
We studied the effects of high mobility group box chromosomal protein 1 (HMGB1) and toll-like receptor (TLR4) in diisonoyl phthalate (DINP)-induced asthma. Mice with DINP-induced asthma were treated with a TLR4-signaling inhibitor or anti-HMGB1 antibody, and various markers of asthma were measured 24 h later. DINP increased airway hyperresponsiveness, numbers of cells in BALF, numbers of inflammatory cells (leukocytes, lymphocytes, monocytes, eosinophils, neutrophils, basophils) in blood, mucus production, pulmonary fibrosis, Th2 type cytokine levels in BALF, and lung cell apoptosis. On the other hand, administrations of TLR4-signaling inhibitors (TAK-242) or anti-HMGB1 antibodies to a mouse model of DINP-induced asthma reduced biological markers of asthma. These results show TLR4 and HMGB1 both contribute to DINP-induced asthma, and that the inhibitions of TLR4 or HMGB1 offer potential means of treating asthma induced by phthalates like DINP.


Cascade hydroarylation/Diels-Alder cycloaddition of alkynylindoles with electron-deficient alkynes and alkenes.
Herein, a novel cascade gold(I)-catalyzed hydroarylation of alkynylindoles and subsequent Diels-Alder cycloaddition with electron-deficient alkynes and alkenes is described. A variety of azepino-fused hydrocarbazoles and carbazoles were obtained in moderate to excellent yields. Key features of this methodology are low catalyst loadings, high regioselectivity, broad functional group tolerances, access to important heterocycles, and 100% atom economy.


Short-term safety of buprenorphine/naloxone in HIV-seronegative opioid-dependent Chinese and Thai drug injectors enrolled in HIV Prevention Trials Network 058.
Buprenorphine/naloxone (BUP/NX) is not licenced for use in China or Thailand and there was little clinical experience with this drug combination in these countries at the inception of HIV Prevention Trial Network (HPTN) 058, a randomized trial comparing risk reduction counselling combined with either short-term or long-term medication assisted treatment with BUP/NX to prevent HIV infection and death amongst opioid-dependent injectors. We conducted a safety phase that included the first 50 subjects enrolled at each of the three initial study sites (N=150). Clinical and laboratory assessments were conducted at baseline and weekly for the first 4 weeks. Changes in laboratory parameters were estimated with random effects models. BUP/NX was well tolerated by study subjects and opioid withdrawal scores decreased substantially during the 3-day induction. Two participants experienced grade 3 clinical adverse events, which were categorized as probably not related to the study drug. Grade 2 or 3 increases in alanine aminotransferase (ALT) occurred in 25 (17%) subjects. The magnitude of ALT increase over 4-week follow-up was strongly associated with baseline ALT elevation. In Chinese and Thai opioid-dependent injectors, we found BUP/NX to be effective in reducing opioid withdrawal symptoms and safe during short-term use. ALT increases were observed over 4-week-follow-up, which are consistent with reports from Western populations. Long-term safety and efficacy evaluations are indicated.


Routine clinical assessment of cognitive functioning in schizophrenia, major depressive disorder, and bipolar disorder.
As more evidence points to the association of cognitive dysfunction with mental health disorders, the assessment of cognitive function in routine clinical care of these disorders is increasingly important. Despite this, it remains unknown how cognitive function is measured in routine clinical practice. The objective of this study was to assess psychiatrists' awareness of cognitive dysfunction in mental health disorders and their methods of cognitive assessment. An online survey was disseminated to psychiatrists in Europe, Asia, Australia and the United States. The survey asked about their perceptions of cognitive dysfunction in several mental health disorders, knowledge of cognitive assessment, method of cognitive assessment, and instruments used to measure cognitive function. Among the 61 respondents, most perceived that schizophrenia was associated with the greatest cognitive dysfunction. Many were unaware whether guidelines were available on cognitive assessment. In schizophrenia, 59% of psychiatrists reportedly used cognitive instruments, while the remainder relied solely on patient history interviews. The use of instruments to assess cognition in major depressive disorder (MDD) and bipolar disorder (BPD) was lower, 38% and 37% respectively. Of the reported instruments used, only a few were actually appropriate for use in the diseases of interest (12% in schizophrenia, 3% in MDD and 0% in BPD). Other instruments reported were clinical measures that did not assess cognition. These findings reveal some inconsistencies in psychiatrists' routine clinical evaluation of cognitive function. There appeared to be low use of true cognitive assessment instruments in clinical practice and confusion regarding what constituted a cognitive assessment instrument.


Detection of the plasmid-mediated mcr-1 gene conferring colistin resistance in human and food isolates of Salmonella enterica and Escherichia coli in England and Wales.
In response to the first report of transmissible colistin resistance mediated by the mcr-1 gene in Escherichia coli and Klebsiella spp. from animals and humans in China, we sought to determine its presence in Enterobacteriaceae isolated in the UK. The PHE archive of whole-genome sequences of isolates from surveillance collections, submissions to reference services and research projects was retrospectively analysed for the presence of mcr-1 using Genefinder. The genetic environment of the gene was also analysed. Rapid screening of the genomes of ∼24 000 Salmonella enterica, E. coli, Klebsiella spp., Enterobacter spp., Campylobacter spp. and Shigella spp. isolated from food or humans identified 15 mcr-1-positive isolates. These comprised: 10 human S. enterica isolates submitted between 2012 and 2015 (8 Salmonella Typhimurium, 1 Salmonella Paratyphi B var Java and 1 Salmonella Virchow) from 10 patients; 3 isolates of E. coli from 2 patients; and 2 isolates of Salmonella Paratyphi B var Java from poultry meat imported from the EU. The mcr-1 gene was located on diverse plasmids belonging to the IncHI2, IncI2 and IncX4 replicon types and its association with ISApl1 varied. Six mcr-1-positive S. enterica isolates were from patients who had recently travelled to Asia. Analysis of WGS data allowed rapid confirmation of the presence of the plasmid-mediated colistin resistance gene mcr-1 in diverse genetic environments and plasmids. It has been present in E. coli and Salmonella spp. harboured by humans in England and Wales since at least 2012.


Oxidative stress after thrombolysis-induced reperfusion in human stroke.
Animal models of transient ischemia suggest that oxygen-derived free radicals produced on reperfusion of ischemic brain could constitute the main cause of reperfusion injury. We aimed to determine the presence and role of lipid peroxidation and protein oxidation-related molecules after tissue plasminogen activator-induced recanalization in human stroke. A total of 160 patients with strokes involving the middle cerebral artery and treated with tissue plasminogen activator and 60 healthy controls were included. Blood samples, transcranial Doppler recordings, and National Institutes of Health Stroke Scale scores were obtained at baseline (pretreatment), 1 hour and 2 hours after tissue plasminogen activator bolus, and 12 hours and 24 hours after stroke onset. The main lipid peroxidation end-product malondialdehyde, advanced oxidation protein products, and plasma concentrations of myeloperoxidase were assessed. At baseline, all oxidative stress biomarkers were higher than in control subjects (P<0.01 for all comparisons). Malondialdehyde remained high compared with controls during the study period, whereas myeloperoxidase concentrations were significantly raised at baseline, 1 hour after tissue plasminogen activator administration, and 12 hours after stroke onset. Malondialdehyde concentrations correlated with stroke severity and were associated with outcome and with hemorrhagic complications. Regarding recanalization, among those patients with middle cerebral artery recanalization by the end of tissue plasminogen activator infusion (44%) or anytime thereafter, no peaking of any of the studied molecules could be identified. Our study showed that systemic oxidative damage to lipids and proteins had already occurred at baseline in stroke. In contrast to animal studies, a relationship between free radical-mediated oxidative damage to lipids or proteins and reperfusion injury after arterial recanalization could not be established.


Reduction of NO adlayers on Pt(110) and Pt(111) in acidic media: evidence for adsorption site-specific reduction.
We present a combined in situ Fourier transform infrared reflection-absorption spectroscopy and voltammetric study of the reduction of saturated and subsaturated NO adlayers on Pt(111) and Pt(110) single-crystal surfaces in acidic media. The stripping voltammetry experiments and the associated evolution of infrared spectra indicate that different features (peaks) observed in the voltammetric profile for the electrochemical reduction of NO adlayers on the surfaces considered are related to the reduction of NO(ads) at different adsorption sites and not to different (consecutive) processes. More specifically, reduction of high- and intermediate-coverage (ca. 0.5-1 monolayers (ML)) NO adlayers on Pt(110) is accompanied by site switching from atop to bridge position, in agreement with the ultra-high-vacuum data. On Pt(111) linearly bonded (atop) NO and face-centered cubic 3-fold-hollow NO species coexist at high coverages (0.25-0.5 ML) and can be reduced consecutively and independently. On Pt(111) and Pt(110) electrodes, linearly bonded NO species are more reactive than multifold-bonded NO species. Both spectroscopic and voltammetric data indicate that ammonia is the main product of NO(ads) reduction on the two surfaces examined.


Particle Monte Carlo simulation of string-like colloidal assembly in two and three dimensions.
We simulate structural phase behavior of polymer-grafted colloidal particles by molecular Monte Carlo technique. The interparticle potential, which has a finite repulsive square-step outside a rigid core of the colloid, was previously confirmed via numerical self-consistent field calculation. This model potential is purely repulsive. We simulate these model colloids in the canonical ensemble in two and three dimensions and find that these particles containing no interparticle attraction self-assemble and align in a string-like assembly, at low temperature and high density. This string-like colloidal assembly is related to percolation phenomena. Analyzing the cluster size distribution and the average string length, we build phase diagrams and discover that the average string length diverges around the region where the melting transition line and the percolation transition line cross. This result is similar to Ising spin systems, in which the percolation transition line and the order-disorder line meet at a critical point.


Anti-inflammatory activities of alpha-truxillic acid derivatives and their monomer components.
The anti-inflammatory activities of alpha-truxillic acid (1) and 4,4'-dihydroxy-alpha-truxillic acid (2) as well as their monomer components (E)-cinnamic acid (3) and (E)-p-coumaric acid (4) were evaluated in the formalin test. alpha-Truxillic acid (1) and its derivative 4,4'-dihydroxy-alpha-truxillic acid (2) exhibited significant activity against inflammatory pain response, while their monomer components (E)-cinnamic acid (3) and (E)-p-coumaric acid (4) did not show any activity against either neurogenic or inflammatory pain responses induced by formalin in mice. These results suggested that the dimeric structure might play an important role for the expression of anti-inflammatory activity. Furthermore, in order to gain information on their potencies, their anti-inflammatory activities were compared with that of incarvillateine (5) which contains the same dimeric structure and showed more potent antinociceptive activity than morphine in the formalin test. The activities of alpha-truxillic acid (1) and 4,4'-dihydroxy-alpha-truxillic acid (2) at the dose of 40 mg/kg against inflammatory pain response were equal to that of incarvillateine at doses of 20 mg/kg.


State-space models of impulse hemodynamic responses over motor, somatosensory, and visual cortices.
THE PAPER PRESENTS STATE SPACE MODELS OF THE HEMODYNAMIC RESPONSE (HR) OF FNIRS TO AN IMPULSE STIMULUS IN THREE BRAIN REGIONS: motor cortex (MC), somatosensory cortex (SC), and visual cortex (VC). Nineteen healthy subjects were examined. For each cortex, three impulse HRs experimentally obtained were averaged. The averaged signal was converted to a state space equation by using the subspace method. The activation peak and the undershoot peak of the oxy-hemoglobin (HbO) in MC are noticeably higher than those in SC and VC. The time-to-peaks of the HbO in three brain regions are almost the same (about 6.76 76 ± 0.2 s). The time to undershoot peak in VC is the largest among three. The HbO decreases in the early stage (~0.46 s) in MC and VC, but it is not so in SC. These findings were well described with the developed state space equations. Another advantage of the proposed method is its easy applicability in generating the expected HR to arbitrary stimuli in an online (or real-time) imaging. Experimental results are demonstrated.


Comparison of Efficiency Between Corticosteroid and Platelet Rich Plasma Injection Therapies in Patients With Knee Osteoarthritis.
This study aims to assess whether platelet rich plasma (PRP) is an effective treatment for knee osteoarthritis, and compare its efficiency with corticosteroid treatment in terms of pain control, physical function, and quality of life. The study included 50 patients (4 males, 46 females; mean age 61.6±6.9 years; range 50 to 75 years) who were diagnosed as grade 3 knee osteoarthritis. Patients were randomized into three groups as corticosteroid group (receiving one corticosteroid injection), single PRP group (receiving one PRP injection), and three PRP group (receiving three PRP injections with one week interval). All patients were given a home exercise program. Patients were evaluated with Visual Numeric Scale (VNS), Western Ontario and McMaster Universities Arthritis Osteoarthritis Index (WOMAC), Lequesne index and the Hospital Anxiety and Depression Scale before treatment, and at second and sixth months following the implementation of injections. In single and three PRP groups; VNS, WOMAC and Lequesne scores decreased significantly at second month follow-up; also, sixth month scores showed a slight increase but remained significantly lower than baseline. In corticosteroid group, all VNS, WOMAC, and Lequesne scores decreased at second month follow-up; however, at sixth month, rest, night VNS and WOMAC stiffness scores were increased while no significant difference was found with baseline. At sixth month; VNS movement, WOMAC pain, function, and the total and Lequesne scores were worse, but remained significantly lower than baseline. When groups were compared, three PRP group's second month VNS movement scores were significantly lower than the corticosteroid group. Sixth month VNS movement and WOMAC pain scores were significantly lower in single and three PRP groups compared to the corticosteroid group, with no significant difference between the PRP groups. Our findings revealed that PRP is a safe treatment option and efficient in osteoarthritis symptom control up to six months after application. Treatment response obtained with corticosteroid injection has a shorter duration than PRP treatment.


Oxidized low-density lipoprotein as a delivery system for photosensitizers: implications for photodynamic therapy of atherosclerosis.
Photodynamic therapy is a promising new strategy in the treatment of cardiovascular diseases. Photodynamic therapy for vascular diseases may be improved by the specific delivery of photosensitizers to the atherosclerotic lesion. In this study, we studied whether oxidatively modified low-density lipoprotein (OxLDL) could be used as a specific carrier for photosensitizers, thereby using the scavenger receptor expressed on macrophages as a target. The photosensitizer aluminum phthalocyanine chloride (AlPc) was incorporated into OxLDL, and its photodynamic effects were studied. Macrophages (RAW 264.7) were incubated with various concentrations of OxLDL-AlPc for different periods. After illumination of the cells with red light, cytotoxicity was observed that was dependent on the time of illumination and incubation. Macrophages incubated with OxLDL-AlPc that were not illuminated revealed no cytotoxicity. The uptake of the OxLDL-AlPc complexes was mediated by scavenger receptors expressed on macrophages. In the presence of the polyanion polyinosinic acid, a specific ligand for scavenger receptors, no cytotoxicity could be observed. Serum incubations of the OxLDL-AlPc complexes revealed that these complexes stay intact after incubation. No redistribution of AlPc to other plasma (lipo-) proteins could be detected, and 80-90% of the AlPc remained associated with the OxLDL particle. These results indicate that OxLDL may function as a specific delivery system for photosensitizers to the scavenger receptors expressed on the macrophages in the atherosclerotic lesion, increasing the beneficial effects of photodynamic therapy for cardiovascular diseases.


The elderly hypertensive: a neglected patient?
A cohort of 1,002 elderly hypertensive patients who received care at six family practice residency program clinics in Iowa was followed for nearly four years in a historical prospective design study. Demographic and clinical data were abstracted and the end points of the study were the occurrence of a cerebrovascular accident (n = 37), myocardial infarction (n = 27), or death (n = 102). Survival regression analysis showed that the risk of cerebrovascular accident is greater than any other major morbid event and that risk is proportional to increasing levels of both systolic and diastolic blood pressure. The findings are in agreement with published studies of hypertension and its treatment in other age groups.


Bullous pemphigoid: using animal models to study the immunopathology.
Bullous pemphigoid was first described by Lever in 1953 as a subepidermal blistering disease. Its immunohistological features include dermal-epidermal junction separation, an inflammatory cell infiltrate in the upper dermis, and basement membrane zone-bound autoantibodies. These autoantibodies show a linear staining at the dermal-epidermal junction, activate complement, and recognize two major hemidesmosomal antigens, BP230 (BPAG1) and BP180 (BPAG2 or type XVII collagen). An IgG passive transfer mouse model of BP was developed by administering rabbit antimurine BP180 antibodies to neonatal mice. This model recapitulates the key features of human bullous pemphigus. Using this in vivo model system, several key cellular and molecular events leading to the bullous pemphigus disease phenotype were identified, including IgG binding, complement activation, mast cell degranulation, and neutrophil infiltration and activation. Proteinases and reactive oxygen species released by neutrophils work together to damage the basement membrane zone, causing dermal-epidermal junction separation. Recent experimental data from human bullous pemphigus studies suggest that human bullous pemphigus and its mouse IgG passive transfer model counterpart may well share not only common immunohistological features but also pathological mechanisms underlying the development of this antibody-mediated disease.


Antibacterial activities of the methanol extracts and compounds from Erythrina sigmoidea against Gram-negative multi-drug resistant phenotypes.
In the present study, the methanol extracts from the leaves, as well as compounds namely sigmoidin I (1), atalantoflavone (2), bidwillon A (3), neocyclomorusin (4), 6α-hydroxyphaseollidin (5) and neobavaisoflavone (6) (from the bark extract) were tested for their activities against a panel of Gram-negative bacteria including multi-drug resistant (MDR) phenotypes. Broth microdilution method was used to determine the minimum inhibitory concentrations (MICs) and the minimum bactericidal concentrations (MBCs) of the extracts as well as compounds 1-6. The MIC results indicated that the crude extracts from the leaves and bark of this plant were able to inhibit the growth of 96.3 % of the 27 tested bacteria. Compounds 2-6 displayed selective activities, their inhibitory effects being obtained on 8.3 %, 41.7 %, 58.3 %, 58.3 % and 66.7 % of tested bacteria respectively for 2, 3, 5, 6 and 4. The lowest MIC value of 8 μg/mL was obtained with 6 against Escherichia coli ATCC8739, Enterobacter cloacae ECCI69, Klebsiella pneumoniae KP55, Providencia stuartii NAE16 and Pseudomonas aeruginosa PA01. The present study demonstrates that Erythrina sigmoidea is a potential source of antibacterial drugs to fight against MDR bacteria. Neobavaisoflavone (6) is the main antibacterial consituents of the bark crude extract.


Immunogenicity of iron-regulated outer membrane proteins of Pasteurella multocida A:3 in cattle: molecular characterization of the immunodominant heme acquisition system receptor (HasR) protein.
The iron-regulated outer membrane proteins (IROMPs) of Pasteurella multocida A:3 strain 232 (Pm232), a bovine isolate, were investigated as potential immunogens in cattle. We addressed the ability of P. multocida IROMP-enriched fractions to induce antibody responses in cattle by different vaccination strategies and the protective efficacy of these antibodies using a P. multocida-induced pneumonia challenge model. Vaccination of cattle with outer membrane-enriched fractions derived from Pm232 grown on either iron-depleted (IROMPs) or iron-sufficient (OMPs) conditions induced significant antibody responses; however, the correlation with lung lesion scores was not significant (P = 0.01 and P < 0.07, respectively). SDS-PAGE, Western blots and densitometric analyses of Pm232 grown under iron-deficient conditions revealed five major IROMPs including an immunodominant 96 kDa protein band. Mass spectrometry analysis of the 96kDa protein band suggested homology with the heme acquisition system receptor (HasR) of avian P. multocida (strain Pm70) and was confirmed by DNA sequence analysis of the cloned Pm232 hasR gene. Further analyses indicated that Pm232 HasR is a surface-exposed OMP and conserved among most P. multocida isolates investigated. In addition, cattle vaccinated with live Pm232 or IROMPs had significantly higher antibody responses to the 96 kDa protein band and the correlation with lung lesion scores approached significance (P = 0.056). These results indicate that antibody responses in cattle are induced by P. multocida IROMPs, and that the 96 kDa HasR protein is an immunodominant IROMP.


QMHC interview: Karl Albrecht. Interview by Marie E. Sinioris.
For over a decade, Karl Albrecht has helped health care and other organizations focus on creating superior value for customers. In his books Service America! and The Only Thing That Matters, he developed a model of organizational improvement called Total Quality Service that is tailored to service organizations. One of the principles underlying this model is that service industries are fundamentally different from manufacturing industries. Thus, the approach to quality improvement in services should not be based on a manufacturing mentality. QMHC interviewed Karl Albrecht to catch up with his latest thinking on value creation, leadership, and innovation in health care and elsewhere.


Polysaccharides of a fermented food, natto, suppress sucrose-induced hyperglycemia in an in vivo evaluation system and inhibit glucose uptake by human intestinal cells.
Natto is a well-known traditional Japanese food produced by fermenting soybeans with Bacillus subtilis var natto. Here we found that the water-soluble viscous fraction of natto inhibits sucrose- or glucose-induced hyperglycemia in silkworms. The water-soluble viscous fraction treated with DNase I, RNase A, and proteinase K, followed by phenol extraction also suppressed sucrose-induced hyperglycemia in silkworms. The enzyme-treated polysaccharide fraction of natto inhibits glucose uptake by Caco-2 cells, human intestinal epithelial cells. These findings suggest that the polysaccharide components of natto selected on the basis of their suppressive effects on sucrose-induced hyperglycemia in silkworms inhibit glucose uptake by human intestinal cells.


Graphene-based electrochemical sensors.
Graphene, one kind of emerging carbon nanomaterial, has attracted increasing attention recently. Due to its fascinating physical and electrochemical properties, graphene as a promising electrode material has been widely used in electrochemical sensing applications. In this review, different approaches for the fabrication of graphene and the preparation of graphene-modified electrodes for electrochemical sensors are introduced. Moreover, recent research results on different graphene-based materials as an electrochemical platform for the detection of various biomolecules and chemicals are reviewed and compared. More electrochemical studies on this novel material should show up in the near future.


Insulin pumps in hospital: a guide for the generalist physician.
An increasing number of people with type 1 diabetes mellitus are using insulin pump therapy, also known as continuous subcutaneous insulin infusion therapy (CSII). Most patients using pumps are safest remaining on CSII if admitted to hospital, unless incapacitated. This review provides the generalist physician with a framework to guide the management of such patients, although one should always seek specialist diabetes advice where available.


PHD3-dependent hydroxylation of HCLK2 promotes the DNA damage response.
The DNA damage response (DDR) is a complex regulatory network that is critical for maintaining genome integrity. Posttranslational modifications are widely used to ensure strict spatiotemporal control of signal flow, but how the DDR responds to environmental cues, such as changes in ambient oxygen tension, remains poorly understood. We found that an essential component of the ATR/CHK1 signaling pathway, the human homolog of the Caenorhabditis elegans biological clock protein CLK-2 (HCLK2), associated with and was hydroxylated by prolyl hydroxylase domain protein 3 (PHD3). HCLK2 hydroxylation was necessary for its interaction with ATR and the subsequent activation of ATR/CHK1/p53. Inhibiting PHD3, either with the pan-hydroxylase inhibitor dimethyloxaloylglycine (DMOG) or through hypoxia, prevented activation of the ATR/CHK1/p53 pathway and decreased apoptosis induced by DNA damage. Consistent with these observations, we found that mice lacking PHD3 were resistant to the effects of ionizing radiation and had decreased thymic apoptosis, a biomarker of genomic integrity. Our identification of HCLK2 as a substrate of PHD3 reveals the mechanism through which hypoxia inhibits the DDR, suggesting hydroxylation of HCLK2 is a potential therapeutic target for regulating the ATR/CHK1/p53 pathway.


Microbiological quality assessment of fresh produce: Potential health risk to children and urgent need for improved food safety in school feeding schemes.
About 388 million school-going children worldwide benefit from school feeding schemes, which make use of fresh produce to prepare meals. Fresh produce including leafy greens and other vegetables were served at 37% and 31% of school feeding programs, respectively, in Africa. This study aimed at assessing the microbiological quality of fresh produce grown onsite or supplied to South African schools that are part of the national school feeding programs that benefit over 9 million school-going children. Coliforms, Escherichia coli, Enterobacteriaceae, and Staphylococcus aureus were enumerated from fresh produce (n = 321) samples. The occurrence of E. coli, Listeria monocytogenes, Salmonella spp., and extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae was determined. Presumptive pathogens were tested for antimicrobial resistance. E. coli was further tested for diarrheagenic virulence genes. Enterobacteriaceae on 62.5% of fresh produce samples (200/321) exceeded previous microbiological guidelines for ready-to-eat food, while 86% (276/321 samples) and 31.6% (101/321 samples) exceeded coliform and E. coli criteria, respectively. A total of 76 Enterobacteriaceae were isolated from fresh produce including E. coli (n = 43), Enterobacter spp. (n = 15), and Klebsiella spp. (n = 18). Extended-spectrum β-lactamase production was confirmed in 11 E. coli, 13 Enterobacter spp., and 17 Klebsiella spp. isolates. No diarrheagenic virulence genes were detected in E. coli isolates. However, multidrug resistance (MDR) was found in 60.5% (26/43) of the E. coli isolates, while all (100%; n = 41) of the confirmed ESBL and AmpC Enterobacteriaceae showed MDR. Our study indicates the reality of the potential health risk that contaminated fresh produce may pose to school-going children, especially with the growing food safety challenges and antimicrobial resistance crisis globally. This also shows that improved food safety approaches to prevent foodborne illness and the spread of foodborne pathogens through the food served by school feeding schemes are necessary.


Exposure to the Epstein-Barr Viral Antigen Latent Membrane Protein 1 Induces Myelin-Reactive Antibodies In Vivo.
Multiple sclerosis (MS) is an autoimmune chronic inflammatory disease of the central nervous system (CNS). Cross-reactivity of neuronal proteins with exogenous antigens is considered one of the possible mechanisms of MS triggering. Previously, we showed that monoclonal myelin basic protein (MBP)-specific antibodies from MS patients cross-react with Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1). In this study, we report that exposure of mice to LMP1 results in induction of myelin-reactive autoantibodies in vivo. We posit that chronic exposure or multiple acute exposures to viral antigen may redirect B cells from production of antiviral antibodies to antibodies, specific to myelin antigen. However, even in inbred animals, which are almost identical in terms of their genomes, such an effect is only observed in 20-50% of animals, indicating that this change occurs by chance, rather than systematically. Cross-immunoprecipitation analysis showed that only part of anti-MBP antibodies from LMP1-immunized mice might simultaneously bind LMP1. In contrast, the majority of anti-LMP1 antibodies from MBP-immunized mice bind MBP. De novo sequencing of anti-LMP1 and anti-MBP antibodies by mass spectrometry demonstrated enhanced clonal diversity in LMP1-immunized mice in comparison with MBP-immunized mice. We suggest that induction of MBP-reactive antibodies in LMP1-immunized mice may be caused by either Follicular dendritic cells (FDCs) or by T cells that are primed by myelin antigens directly in CNS. Our findings help to elucidate the still enigmatic link between EBV infection and MS development, suggesting that myelin-reactive antibodies raised as a response toward EBV protein LMP1 are not truly cross-reactive but are primarily caused by epitope spreading.


Liraglutide versus glimepiride monotherapy for type 2 diabetes (LEAD-3 Mono): a randomised, 52-week, phase III, double-blind, parallel-treatment trial.
New treatments for type 2 diabetes mellitus are needed to retain insulin-glucose coupling and lower the risk of weight gain and hypoglycaemia. We aimed to investigate the safety and efficacy of liraglutide as monotherapy for this disorder. In a double-blind, double-dummy, active-control, parallel-group study, 746 patients with early type 2 diabetes were randomly assigned to once daily liraglutide (1.2 mg [n=251] or 1.8 mg [n=247]) or glimepiride 8 mg (n=248) for 52 weeks. The primary outcome was change in proportion of glycosylated haemoglobin (HbA(1c)). Analysis was done by intention-to-treat. This trial is registered with ClinicalTrials.gov, number NTC00294723. At 52 weeks, HbA(1c) decreased by 0.51% (SD 1.20%) with glimepiride, compared with 0.84% (1.23%) with liraglutide 1.2 mg (difference -0.33%; 95% CI -0.53 to -0.13, p=0.0014) and 1.14% (1.24%) with liraglutide 1.8 mg (-0.62; -0.83 to -0.42, p<0.0001). Five patients in the liraglutide 1.2 mg, and one in 1.8 mg groups discontinued treatment because of vomiting, whereas none in the glimepiride group did so. Liraglutide is safe and effective as initial pharmacological therapy for type 2 diabetes mellitus and leads to greater reductions in HbA(1c), weight, hypoglycaemia, and blood pressure than does glimepiride.


Canopy mortality has doubled in Europe's temperate forests over the last three decades.
Mortality is a key indicator of forest health, and increasing mortality can serve as bellwether for the impacts of global change on forest ecosystems. Here we analyze trends in forest canopy mortality between 1984 and 2016 over more than 30 Mill. ha of temperate forests in Europe, based on a unique dataset of 24,000 visually interpreted spectral trajectories from the Landsat archive. On average, 0.79% of the forest area was affected by natural or human-induced mortality annually. Canopy mortality increased by +2.40% year-1, doubling the forest area affected by mortality since 1984. Areas experiencing low-severity mortality increased more strongly than areas affected by stand-replacing mortality events. Changes in climate and land-use are likely causes of large-scale forest mortality increase. Our findings reveal profound changes in recent forest dynamics with important implications for carbon storage and biodiversity conservation, highlighting the importance of improved monitoring of forest mortality.


Solid and cystic tumor of the pancreas with massive hemoperitoneum.
A case of a solid cystic tumor (SCT) of the pancreas with massive hemoperitoneum occurring in a 31-year-old woman is reported. She was admitted with abdominal pain and rebound tenderness. Computed tomography (CT) disclosed a large tumor consisting of both solid and cystic components in the body of the pancreas. Within 12 hours after admission, she exhibited exacerbation of pain and hemoperitoneum on CT with deterioration of general condition and anemia. Celiac artery angiography demonstrated the tumor as a hypovascular mass. SCT of the pancreas was suspected, and emergent laparotomy was performed. In the surgical procedure, tumor in the body of the pancreas with rupture was detected. Distal pancreatectomy was performed with removal of the regional lymph nodes. There was no evidence of metastatic disease in the abdomen. The cut surface of the tumor exhibited areas of cystic degeneration containing hemorrhagic friable materials. Microscopically, the tumor cells were mainly cubic or cylindrical peripherally in pseudo-papillary formation. Postoperative recovery was normal and she was discharged on the 15th postoperative day. Three years after operation, she is free of disease without findings of recurrence.


Immediate endoscopic diagnosis of upper gastrointestinal bleeding. Its accuracy and value in relation to associated pathology.
Two hundred sixty-two patients with active upper gastrointestinal (GI) bleeding underwent panendoscopy between July 1970 and March 1973. There was 100% accuracy of endoscopic diagnosis as to the anatomical site of bleeding; the etiopathologic definition was 94.7% accurate. The series was divided into two groups, 116 with "liver disease" and 146 with "no liver disease." There were 107 patients with varices: 21 fell into no liver disease (small varices) and 86 into liver disease (39 small and 47 large varices). All had associated gastritis. Three endoscopic bleeding patterns were identified in the liver disease group. Only 27% of the patients in the liver disease group with varices (cirrhotics) had frank variceal hemorrhage, whereas 57% bled from hemorrhagic gastritis. The diagnostic unit provided early diagnosis, meaningful therapy, organized data gathering, and rough estimates of ultimate prognosis.


Profile of head and neck cancers in Dr. Lütfi Kırdar Kartal Educational and Research Hospital.
Head and neck cancers account for 17.6% of all malignant neoplasms. Data on the incidence of head and neck cancers and histological subtypes in Turkey are limited. This study aimed to provide an overview of statistics for head and neck cancers. This retrospective and descriptive study included 1973 cases of in-situ carcinomas, malignant and borderline neoplasia from the head and neck region in a period of 5 years, diagnosed at Dr.Lütfi Kırdar Kartal Education and Research Hospital's Department of Pathology. Demographic data of patients and data from histopathological report were recorded. The topography of all tumours was coded according to the International Classification of Diseases for Oncology, 2nd and 3rd edition and histological and behavioral codes were given according to ICD-O-3. Data was entered and analyzed using Microsoft Excel™, version 2010. Male and female ratio was 1.03 for head and neck cancers. The most common sites for males were described as the ratio in head and neck cancers and all malignant tumours as follows: the skin (non-melanoma) (48.4%-8.2%), the larynx (12.6%-2.3%), the thyroid (11.1%-1.2%), the lips (7.8%-1.3%), and the nasopharynx (3.1%-0.5%). The most common sites for females were as follows: the thyroid (45.3%-8.3%), the skin (non-melanoma) (39.6%-7.3%), the lips (3.6%-0.6%), the nasopharynx (2.2%-0.4%) and the eye (1.7%-0.3%). The male: female ratio was 25.2 for larynx cancers. The most common histological types and rates for males and females were also given. This descriptive epidemiological pathology-based cancer incidence study has relative value for describing head and neck cancer incidences in the İstanbul region and shown that pathology report-based descriptive epidemiological studies are still valuable for determination of the regional distribution of cancer incidence out of the active cancer registration regions.


Intracoronary brachytherapy. Treatment of in-stent restenosis with the Beta-Cath system. Initial experience in Latin America.
To assess the safety and efficacy of intracoronary brachytherapy using the Beta-Cath system for preventing recurrence of in-stent restenosis (ISR), by analyzing clinical, angiographic, and intracoronary ultrasound (ICUS) results. This study assessed 30 patients with ISR in native coronary arteries who underwent balloon catheter angioplasty followed by intracoronary beta radiation with the Beta-Cath system (90Sr/Y). The study comprised complex, extensive (18.66 +/- 4.15 mm) restenotic lesions, 77% of which were of the diffuse-proliferative type. Brachytherapy was successful in 100% of the cases. The mean radiation dose used was 20.7 +/- 2.3 Gy, released for a mean period of 3.8 +/- 2.1 minutes. On late follow-up, the in-stent minimum luminal diameter (MLD) slightly decreased (from 1.98 +/- 0.30 mm to 1.84 +/- 0.39 mm at 6 months; P=0.13), with a late loss of 0.14 +/- 0.18 mm. The intrasegmentary MLD was significantly smaller than the in-stent diameter (1.55 +/- 0.40 mm vs 1.84 +/- 0.39 mm; P=0.008), and was associated with a more significant late loss (0.40 +/- 0.29 mm vs 0.14 +/- 0.18 mm; P=0.0001). On ICUS, a mild increase of 6.8 +/- 14.3 mm3 in the neointimal tissue was observed at 6 months (P=0.19), and the percentage of volumetric obstruction increased by 4.7 +/- 7.5%. Binary restenosis and revascularization of the target vessel recurred in 17% of the cases; late occlusion associated with myocardial infarction occurred in 1 case (3%). Event-free survival was 80%. The management of in-stent restenosis with intracoronary beta radiation proved to be a safe and effective procedure, with a high rate of immediate success, representing a therapeutic option for inhibiting neointimal hyperplasia.


Validation of a murine proteome-wide phage display library for identification of autoantibody specificities.
Autoimmunity is characterized by loss of tolerance to tissue-specific as well as systemic antigens, resulting in complex autoantibody landscapes. Here, we introduce and extensively validate the performance characteristics of a murine proteome-wide library for phage display immunoprecipitation and sequencing (PhIP-seq) in profiling mouse autoantibodies. This library was validated using 7 genetically distinct mouse lines across a spectrum of autoreactivity. Mice deficient in antibody production (Rag2-/- and μMT) were used to model nonspecific peptide enrichments, while cross-reactivity was evaluated using anti-ovalbumin B cell receptor-restricted OB1 mice as a proof of principle. The PhIP-seq approach was then utilized to interrogate 3 distinct autoimmune disease models. First, serum from Lyn-/- IgD+/- mice with lupus-like disease was used to identify nuclear and apoptotic bleb reactivities. Second, serum from nonobese diabetic (NOD) mice, a polygenic model of pancreas-specific autoimmunity, was enriched in peptides derived from both insulin and predicted pancreatic proteins. Lastly, Aire-/- mouse sera were used to identify numerous autoantigens, many of which were also observed in previous studies of humans with autoimmune polyendocrinopathy syndrome type 1 carrying recessive mutations in AIRE. These experiments support the use of murine proteome-wide PhIP-seq for antigenic profiling and autoantibody discovery, which may be employed to study a range of immune perturbations in mouse models of autoimmunity profiling.


Who is at risk for having persistent insomnia symptoms? A longitudinal study in the general population in Korea.
Our study had three goals: (1) to investigate the longitudinal course of insomnia symptoms over 4 years (3 time points) by analyzing the trajectory of insomnia symptoms for all participants, (2) to compare persistent insomnia symptom to nonpersistent insomnia symptom groups on mental health and quality of life (QoL), and (3) to conduct exploratory analyses on the relative contribution of multiple factors to persistence of insomnia symptoms. Our population-based longitudinal study utilized a community-based sample from the Korean Genome and Epidemiology study (KoGES). Participants were 1247 individuals (40.1% men; mean age, 54.3±7.1 years). Insomnia, QoL (measured by 12-item Short-Form health survey [SF-12]), sleep-interfering behaviors, and depression (measured by the Beck Depression Inventory [BDI]) were followed with biennial examinations at 3 data points spaced 2 years apart (baseline, time 1, and time 2). Among individuals experiencing insomnia symptoms at baseline, the most common trajectory was to experience persistent nocturnal insomnia symptoms across all 3 time points. Those with persistent insomnia symptoms had significantly lower physical and mental QoL (measured by SF-12) and higher depression (measured by BDI) at time points compared to those without persistent nocturnal insomnia symptoms. A follow-up exploratory receiver operating characteristic curve (ROC) analysis identified poor sleep quality, frequent sleep-interfering behaviors, and low mental health QoL as the strongest predictors of persistent insomnia symptoms above other well-known risk factors. In particular, an interaction between poor sleep quality, sleep-interfering behaviors, and mental health QoL appeared to be the strongest risk factor for persistent insomnia symptoms.


Quality of life (QOL) in patients with acromegaly is severely impaired: use of a novel measure of QOL: acromegaly quality of life questionnaire.
Acromegaly Quality of Life Questionnaire (AcroQoL) is a new disease-generated quality of life (QOL) questionnaire comprising 22 questions covering physical and psychological aspects of acromegaly and subdivided into "appearance" and "personal relations" categories. We have performed a cross-sectional study of QOL in 80 patients [43 male (mean age, 54.2 yr; range, 20-84); median GH, 0.93ng/ml (range, <0.3 to 23.7); IGF-I, 333.1 ng/ml (range, 47.7-899)] with acromegaly. In addition to AcroQoL, patients completed three generic QOL questionnaires: Psychological General Well-Being Schedule (PGWBS), EuroQol, and a signs and symptoms score (SSS). All three generic questionnaires confirmed impairment in QOL [mean scores: PGWBS, 69.6; EuroQol, visual analog scale, 66.4 (range, 20-100) and utility index, 0.7 (range, -0.07 to 0.92); and SSS, 12 (range, 0-27)]. There was no correlation between biochemical control and any measure of QOL. AcroQoL (57.3%; range, 18.2-93.2) correlated with PGWBS (r = 0.73; P < 0.0001); and in patients with active disease, AcroQoL-physical dimension correlated with SSS (r = -0.67; P < 0.0003). In all questionnaires, prior radiotherapy was associated with impaired QOL. In conclusion, these data underline the marked impact that acromegaly has on patients' QOL and provide the first evidence validating AcroQoL against well-authenticated measures of QOL. This indicates the potential of AcroQoL as a patient-friendly measure of disease activity.


Conformations of oxidized cytochrome c oxidase.
Oxidized cytochrome c oxidase is shown to exist in three conformations in addition to the transient "g5" conformation previously reported [Shaw, R. W., Hansen, R. E., & Beinert, H. (1978) J. Biol. Chem. 253, 6637-6640]. The "resting" and "g12" conformations are distinguished by an NO-induced cytochrome a3 electron paramagnetic resonance (EPR) signal and an EPR signal at g' = 12, respectively. The "oxygenated" conformatin exhibits an unusual EPR signal in the presence of fluoride and is identical with the "oxygenated" state first discovered by Okunuki et al. [Okunuki, K., Hagihora, B., Sekuzu, I., & Horio, T. (1958) Proc. Int. Symp. Enzyme Chem., Tokyo, Kyoto, 264]. It is proposed that when the reduced enzyme is reoxidized by dioxygen, the oxidized enzyme first relaxes from the "g5" into the "oxygenated" conformation after which a percentage of the molecules slowly relax into the "g12" conformation. The "resting" conformation is not formed when the enzyme is reoxidized. On the basis of the EPR observations, it is proposed that these various conformations of the oxidized enzyme differ in the structure of the cytochrome a3--Cua3 site. Structures for the cytochrome a3--Cua3 site are proposed for each conformation, and a mechanism by which these conformations undergo interconversion among themselves is described.


Are students ready for meaningful use?
The meaningful use (MU) of electronic medical records (EMRs) is being implemented in three stages. Key objectives of stage one include electronic analysis of data entered into structured fields, using decision-support tools (e.g., checking drug-drug interactions [DDI]) and electronic information exchange. The authors assessed the performance of medical students on 10 stage-one MU tasks and measured the correlation between students' MU performance and subsequent end-of-clerkship professionalism assessments and their grades on an end-of-year objective structured clinical examination. Two-hundred and twenty-two third-year medical students on the internal medicine (IM) clerkship. DESIGN/MAIN MEASURES: From July 2010 to February 2012, all students viewed 15 online tutorials covering MU competencies. The authors measured student MU documentation and performance in the chart of a virtual patient using a fully functional training EMR. Specific MU measurements included, adding: a new problem, a new medication, an advanced directive, smoking status, the results of screening tests; and performing a DDI (in which a major interaction was probable), and communicating a plan for this interaction. A total of 130 MU errors were identified. Sixty-eight (30.6%) students had at least one error, and 30 (13.5%) had more than one (range 2-6). Of the 130 errors, 90 (69.2%) were errors in structured data entry. Errors occurred in medication dosing and instructions (18%), DDI identification (12%), documenting smoking status (15%), and colonoscopy results (23%). Students with MU errors demonstrated poorer performance on end-of-clerkship professionalism assessments (r =-0.112, p=0.048) and lower observed structured clinical examination (OSCE) history-taking skills (r =-0.165, p=0.008) and communication scores (r= - 0.173, p=0.006). MU errors among medical students are common and correlate with subsequent poor performance in multiple educational domains. These results indicate that without assessment and feedback, a substantial minority of students may not be ready to progress to more advanced MU tasks.


Cholesterol Prevents Hypoxia-Induced Hypoglycemia by Regulation of a Metabolic Ketogenic Shift.
Blood cholesterol levels have been connected to high-altitude adaptation. In the present study, we treated mice with high-cholesterol diets following exposure to acute hypoxic stress and evaluated the effects of the diets on whole-body, liver glucose, and liver fat metabolism. For rapid cholesterol liver uptake, 6-week-old male C57BL/J6 mice were fed with high-cholesterol/cholic acid (CH) diet for 6 weeks and then were exposed to gradual oxygen level reduction for 1 h and hypoxia at 7% oxygen for additional 1 hour using a hypoxic chamber. Animals were than sacrificed, and metabolic markers were evaluated. Hypoxic treatment had a strong hypoglycemic effect that was completely blunted by CH treatment. Decreases in gluconeogenesis and glycogenolysis as well as an increase in ketone body formation were observed. Such changes indicate a metabolic shift from glucose to fat utilization due to activation of the inducible nitric oxide synthase/AMPK axis in the CH-treated animals. Increased ketogenesis was also observed in vitro in hepatocytes after cholesterol treatment. In conclusion, our results show for the first time that cholesterol contributes to metabolic shift and adaptation to hypoxia in vivo and in vitro through induction of HIF-1α and iNOS expression.


Comparative whole genome analysis of dengue virus serotype-2 strains differing in trans-endothelial cell leakage induction in vitro.
The role of genetic differences among dengue virus (DENV) in causing increased microvascular permeability is less explored. In the present study, we compared two closely related DENV serotype-2 strains of Cosmopolitan genotype for their in vitro infectivity phenotype and ability to induce trans-endothelial leakage. We found that these laboratory strains differed significantly in infecting human microvascular endothelial cells (HMEC-1) and hepatocytes (Huh7), two major target cells of DENV in in vivo infections. There was a reciprocal correlation in infectivity and vascular leakage induced by these strains, with the less infective strain inducing more trans-endothelial cell leakage in HMEC-1 monolayer upon infection. The cells infected with the strain capable of inducing more permeability were found to secrete more Non-Structural protein (sNS1) into the culture supernatant. A whole genome analysis revealed 37 predicted amino acid changes and changes in the secondary structure of 3' non-translated region between the strains. But none of these changes involved the signal sequence coded by the C-terminal of the Envelope protein and the two glycosylation sites within the NS1 protein critical for its secretion, and the N-terminal NS2A sequence important for surface targeting of NS1. The strain that secreted lower levels of NS1 and caused less leakage had two mutations within the NS1 protein coding region, F103S and T146I that significantly changed amino acid properties. A comparison of the sequences of the two strains with published sequences of various DENV strains known to cause clinically severe dengue identified a number of amino acid changes which could be implicated as possible key genetic differences. Our data supports the earlier observations that the vascular leakage induction potential of DENV strains is linked to the sNS1 levels. The results also indicate that viral genetic determinants, especially the mutations within the NS1 coding region, could affect this critical phenotype of DENV strains.


Identification of MUC5B mucin gene in human middle ear with chronic otitis media.
To identify the mucin gene and its expressing cells in the middle ear mucosa with chronic otitis media (COM), and to study the correlation between infiltration of inflammatory cells in the submucosa and expression of the mucin gene in the mucosal epithelium with COM. Middle ear mucosal specimens removed from the inferior promontory area of 19 patients undergoing middle ear surgery for COM were studied. Sections were stained with H&E, Alcian blue-periodic acid Schiff (AB-PAS), polyclonal MUC5B antibody, and specific MUC5B riboprobe for histological, histochemical, immunohistochemical, and mucin mRNA analyses. H&E staining revealed pseudostratified epithelia in 18 of the middle ear specimens with COM and cuboidal secretory epithelia in one. AB-PAS staining of epithelia revealed abundant secretory cells and their products (glycoconjugates). In situ hybridization and immunohistochemistry studies demonstrated that the secretory cells of the middle ear mucosa with COM expressed MUC5B mucin mRNA and its product MUC5B mucin. The MUC5B mucin gene and its product were identified in the middle ear secretory cells of patients with COM. Its expression was extensive in pseudostratified mucosal epithelia and related to infiltration of inflammatory cells in the submucosa of the middle ear cleft with COM, suggestive that inflammatory cell products are involved in the production of MUC5B.


The Diversity Outbred Mouse Population Is an Improved Animal Model of Vaccination against Tuberculosis That Reflects Heterogeneity of Protection.
Many studies of Mycobacterium tuberculosis infection and immunity have used mouse models. However, outcomes of vaccination and challenge with M. tuberculosis in inbred mouse strains do not reflect the full range of outcomes seen in people. Previous studies indicated that the novel Diversity Outbred (DO) mouse population exhibited a spectrum of outcomes after primary aerosol infection with M. tuberculosis Here, we demonstrate the value of this novel mouse population for studies of vaccination against M. tuberculosis aerosol challenge. Using the only currently licensed tuberculosis vaccine, we found that the DO population readily controlled systemic Mycobacterium bovis BCG bacterial burdens and that BCG vaccination significantly improved survival across the DO population upon challenge with M. tuberculosis Many individual DO mice that were vaccinated with BCG and then challenged with M. tuberculosis exhibited low bacterial burdens, low or even no systemic dissemination, little weight loss, and only minor lung pathology. In contrast, some BCG-vaccinated DO mice progressed quickly to fulminant disease upon M. tuberculosis challenge. Across the population, most of these disease parameters were at most modestly correlated with each other and were often discordant. This result suggests the need for a multiparameter metric to better characterize "disease" and "protection," with closer similarity to the complex case definitions used in people. Taken together, these results demonstrate that DO mice provide a novel small-animal model of vaccination against tuberculosis that better reflects the wide spectrum of outcomes seen in people.IMPORTANCE We vaccinated the Diversity Outbred (DO) population of mice with BCG, the only vaccine currently used to protect against tuberculosis, and then challenged them with M. tuberculosis by aerosol. We found that the BCG-vaccinated DO mouse population exhibited a wide range of outcomes, in which outcomes in individual mice ranged from minimal respiratory or systemic disease to fulminant disease and death. The breadth of these outcomes appears similar to the range seen in people, indicating that DO mice may serve as an improved small-animal model to study tuberculosis infection and immunity. Moreover, sophisticated tools are available for the use of these mice to map genes contributing to control of vaccination. Thus, the present studies provided an important new tool in the fight against tuberculosis.


Health effects of uranium: new research findings.
Recent plans for a nuclear renaissance in both established and emerging economies have prompted increased interest in uranium mining. With the potential for more uranium mining worldwide and a growth in the literature on the toxicology and epidemiology of uranium and uranium mining, we found it timely to review the current state of knowledge. Here, we present a review of the health effects of uranium mining, with an emphasis on newer findings (2005-2011). Uranium mining can contaminate air, water, and soil. The chemical toxicity of the metal constitutes the primary environmental health hazard, with the radioactivity of uranium a secondary concern. The update of the toxicologic evidence on uranium adds to the established findings regarding nephrotoxicity, genotoxicity, and developmental defects. Additional novel toxicologic findings, including some at the molecular level, are now emerging that raise the biological plausibility of adverse effects on the brain, on reproduction, including estrogenic effects, on gene expression, and on uranium metabolism. Historically, most epidemiology on uranium mining has focused on mine workers and radon exposure. Although that situation is still overwhelmingly true, a smaller emerging literature has begun to form around environmental exposure in residential areas near uranium mining and processing facilities. We present and critique such studies. Clearly, more epidemiologic research is needed to contribute to causal inference. As much damage is irreversible, and possibly cumulative, present efforts must be vigorous to limit environmental uranium contamination and exposure.


Site-specific chemokine expression regulates central nervous system inflammation and determines clinical phenotype in autoimmune encephalomyelitis.
The adoptive transfer of myelin-reactive T cells into wild-type hosts results in spinal cord inflammation and ascending paralysis, referred to as conventional experimental autoimmune encephalomyelitis (EAE), as opposed to brainstem inflammation and ataxia, which characterize disease in IFN-γRKO hosts (atypical EAE). In this article, we show that atypical EAE correlates with preferential upregulation of CXCL2 in the brainstem, and is driven by CXCR2-dependent recruitment of neutrophils. In contrast, conventional EAE is associated with upregulation of CCL2 in the spinal cord, and is driven by recruitment of monocytes via a partially CCR2-dependent pathway. This study illustrates how regional differences in chemokine expression within a target organ shape the spatial pattern and composition of autoimmune infiltrates, leading to disparate clinical outcomes.


Palladium catalyzed diaryl sulfoxide generation from aryl benzyl sulfoxides and aryl chlorides.
Diaryl sulfoxides are synthesized from aryl benzyl sulfoxides and aryl chlorides via three sequential catalytic cycles all promoted by a NiXantPhos-based palladium catalyst. The key step is S-arylation of a sulfenate anion. An air- and moisture-stable precatalyst derived from NiXantPhos efficiently facilitates the transformation. Various functional groups, including those with acidic protons, were tolerated. This method can also be extended to methyl and dibenzyl sulfoxides substrates.


Trefoil factor family mRNA and protein expression in pterygium.
Trefoil factor family (TFF) of proteins are involved in mucosal protection and healing and are induced in inflammatory diseases and neoplastic progression. The purpose of this investigation was to determine if expression of the trefoil factor family (TFF) proteins is altered in human pterygium compared to in normal conjunctiva. Fourteen pterygia (P) and 21 biopsies from normal human conjunctiva (NC) were studied. TFF1, TFF2 and TFF3 mRNA levels were measured by semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR), and TFF1 mRNA levels in addition by real-time PCR. The cellular expression of TFF1 (pS2), TFF3 (intestinal trefoil factor) and M1/MUC5AC mucin in ten pterygia and ten normal human conjunctiva specimens was analyzed by immunohistochemistry using specific monoclonal antibodies. TFF1 mRNA levels were higher in P than in NC (p=0.02). Accordingly, intensity of TFF1 and mucin MUC5AC immunostaining was higher in P than in NC. Mucus-secreting goblet cells (GC) were more densely packed in P than in NC. In both cases, TFF1 protein was detected in GC only, but was not systematically expressed in all GC. In addition, TFF3 mRNA levels were similar (p=0.89) in NC and P, while TFF2 (spasmolytic polypeptide) mRNA were not detected. Both TFF3 and MUC5AC proteins were clearly detected in all GC identified in NC and P. Increased expression of TFF1 mRNA and protein is observed in pterygium GC, suggesting that this trefoil protein might exert protective and beneficial roles during the pathogenesis of this benign and inflammatory conjunctival tumor.


Renal Subcapsular Hematoma Formation Due to Hydronephrosis Caused by Recurrent Uterine Cervical Cancer.
Most non-traumatic renal subcapsular hematomas are found in the presence of primary or metastatic renal tumors, or in the presence of vascular disease of the renal blood vessels. We managed an asymptomatic renal subcapsular hematoma that formed due to uterine cervical cancer that metastasized to the left common iliac lymph nodes. A 48-year-old woman with stage IB1 cervical cancer underwent neoadjuvant chemotherapy and concurrent chemoradiation following a radical hysterectomy. Six months after the completion of her first treatment, she developed left-sided hydronephrosis, a left subcapsular hematoma and left common iliac lymph nodes enlargement as demonstrated with contrast-enhanced computed tomography. Although a renal subcapsular hematoma is rarely a symptom of cervical cancer recurrence, it should be considered if other neoplastic or vascular diseases are ruled out.


Chemiluminometric measurement of atmospheric ozone with photoactivated chromotropic acid.
A highly sensitive, robust, fast, affordable measurement system based on interfacial gas-liquid chemiluminescence (CL) on a wetted transparent screen directly on top of a miniature photomultiplier tube provides the basis of an attractive method for ozone (O(3)). Alkaline chromotropic acid (CA, 4,5-dihydroxynaphthalene-2,7-disulfonic acid) chemiluminesces upon exposure to ozone. No light emission is observed from exposure of alkaline CA to NO(2) or H(2)O(2). However, response to ozone is highly dependent on the age and storage condition of the CA solution. As such, quantitative analysis will require frequent calibration, and the method will not be attractive. We have discovered that photoactivation plays the key role in producing (a) compound(s) from chromotropic acid that appear(s) to be the primary agent(s) responsible for the CL reaction with O(3). We thus devised a method wherein a flowing solution of CA (that is stable in neutral/acidic solutions) is rendered alkaline and then exposed for a few seconds on-line to UV radiation. The solution then reacts with ozone on a screen consisting of an "invisible" nylon stocking that provides for low liquid residence time and high light throughput and results in an LOD of 40 pptv, a determination range at least up to 230 ppbv, and 10-90% and 90-10% response times of 130 and 80 ms, respectively. Intra- and interday repeatabilities at the same concentration were 0.32 and 3.8% in relative standard deviation. On the basis of aging, CL, chromatography, and chromatography-mass spectrometry studies, we suggest that the primary CL-active species are likely dimeric semiquinone species derived from CA by a series of radical reactions.


Blurring boundaries. Interviews with PGT couples about comprehensive chromosome screening.
Comprehensive chromosome examination is a promising approach to Preimplantation Genetic Testing (PGT). Next to testing of specific chromosomes, such as in the case of reduced fertility due to chromosomal translocations, it allows testing of all chromosomes. Hence it potentially reduces the time to pregnancy and the risk of miscarriage. But comprehensive testing also introduces some ethical issues. For example, what is the role of the professional in the decision making regarding embryos with chromosomal abnormalities that are potentially viable? Which chromosomal abnormalities should be communicated to people undergoing fertility treatment? With this paper we wanted to explore the ethical issues related to comprehensive chromosome screening in Preimplantation Genetic Testing. In order to explore these issues, we interviewed seven couples undergoing PGT for chromosomal translocations at the VUB University Hospital, Belgium. We presented them with three fictional cases: the transfer of an embryo with trisomy 21, of an embryo with a sex chromosome aneuploidy and of an embryo with a chromosomal microdeletion. We found that opinions regarding the role of fertility professionals in deciding which embryos to transfer were mixed. Moreover, where to draw the line between healthy and unhealthy embryos was unclear. We also found that couples, although they thought that comprehensive chromosome testing had certain benefits, also considered the increased waiting time for transfer a heavy burden. In the light of comprehensive chromosome screening of embryos, persons undergoing fertility treatment may have views on the burdens and benefits of the techniques that are not analogous to the views of professionals.


Human Dirofilariasis in Bulgaria Between 2009 and 2018.
Autochthonous Dirofilaria repens infections have been found in dogs and other carnivores in most European countries. In the same countries, reports of human dirofilariasis are becoming increasingly prevalent. We present 18 cases of people infected with D. repens for a 10-year period traced in our hospital. The data was collected from the observations and tests of all 18 patients from the whole country treated at the Specialized Hospital for Infectious and Parasitic Diseases in Sofia in the period 2009 - 2018. We used a morphological method, serology test and Knott's method for microfilariae. The patients were 11 to 74 years of age, 12 female and 6 male. In most cases, patients have subcutaneous nodules or face, eyelid and eyes localization. The trend of increasing incidence in Bulgaria continues, with age and sex distribution and localization of the larva being similar to those in other European countries.


[Organization of the substructure of the receptive fields of neurons of the lateral suprasylvian area of the cat cortex].
The substructure of receptive fields of the lateral suprasylvian area neurons was investigated in cats. It was shown that the majority of receptive fields investigated were organized of subelements with different qualitative characteristics according to their responses to moving visual stimuli. With the unmasking method of stimulation, small amplitudes of the motion evoked, as a rule, directional responses, whereas with the masking method the same amplitude of the motion produced nondirectional responses. Some receptive fields of the investigated neurons responded vigorously to the motion of borders of the visual stimuli. The heterogeneity in organization of the substructure of the receptive fields was explained by the convergence of different inputs to the investigated neurons.


Database screening for HIV protease ligands: the influence of binding-site conformation and representation on ligand selectivity.
Screening for potential ligands and docking them into the binding sites of proteins is one of the main tasks in computer-aided drug design. Despite the progress in computational power, it remains infeasible to model all the factors involved in molecular recognition, especially when screening databases of more than 100,000 compounds. While ligand flexibility is considered in most approaches, the model of the binding site is rather simplistic, with neither solvation nor induced complementary usually taken into consideration. We present results for screening different databases for HIV-1 protease ligands with our tool Slide, and investigate the extent to which binding-site conformation, solvation, and template representation generate bias. The results suggest a strategy for selecting the optimal binding-site conformation, for cases in which more than one independent structure is available, and selecting a representation of that binding site that yields reproducible results and the identification of known ligands.