In vitro release studies of furosemide reference tablets: influence of agitation rate, USP apparatus, and dissolution media.
Furosemide is a diuretic drug widely used in chronic renal failure. The drug has low solubility and permeability, which cause clinical problems. Studying the in vitro release performance elucidates the rate and extent of drug dissolved from dosage forms under different conditions. Furosemide reference tablets were tested using USP Apparatuses 1 and 2 as well as the flow-through cell method (USP Apparatus 4), a dissolution apparatus that simulates the human gastrointestinal tract better than the other methods. Dissolution profiles were created with USP Apparatuses 1 and 2 at 25, 50, and 75 rpm and 900 mL of 0.1 M hydrochloric acid, acetate buffer (pH 4.5), and phosphate buffer (pH 6.8). USP Apparatus 4 with a laminar flow of 16 mL/min and 22.6 mm cells was used. Drug dissolution was quantified at 274 nm for 60 min. Mean dissolution time, dissolution efficiency, time to 50% dissolution, and time to 80% dissolution data were used to compare dissolution profiles. Additionally, zero-order, first-order, Higuchi, Hixson-Crowell, Makoid-Banakar, and Weibull models were used to adjust furosemide dissolution data. Between USP Apparatus 1 and 2, significant differences were observed in almost all parameters at 50 and 75 rpm (p < 0.05). A similar dissolution profile (f2 > 50) with a pharmacopoeial dissolution method (USP Apparatus 2 at 50 rpm and 900 mL of phosphate buffer pH 5.8) and USP Apparatus 4 (laminar flow of 16 mL/min, 22.6 mm cells, and pH 6.8) was observed. The Weibull function was the best mathematical model to describe the in vitro release performance of furosemide in the three USP dissolution apparatuses. These results could be used to manufacture better furosemide dosage forms and decrease the negative clinical impact of current furosemide formulations.


A Phase I Clinical Trial with Ex Vivo Expanded Recipient Regulatory T cells in Living Donor Kidney Transplants.
There is considerable interest in therapeutic transfer of regulatory T cells (Tregs) for controlling aberrant immune responses. Initial clinical trials have shown the safety of Tregs in hematopoietic stem cell transplant recipients and subjects with juvenile diabetes. Our hypothesis is that infusion(s) of Tregs may induce transplant tolerance thus avoiding long-term use of toxic immunosuppressive agents that cause increased morbidity/mortality. Towards testing our hypothesis, we conducted a phase I dose escalation safety trial infusing billions of ex vivo expanded recipient polyclonal Tregs into living donor kidney transplant recipients. Despite variability in recipient's renal disease, our expansion protocol produced Tregs which met all release criteria, expressing >98% CD4+CD25+ with <1% CD8+ and CD19+ contamination. Our product displayed >80% FOXP3 expression with stable demethylation in the FOXP3 promoter. Functionally, expanded Tregs potently suppressed allogeneic responses and induced the generation of new Tregs in the recipient's allo-responders in vitro. Within recipients, expanded Tregs amplified circulating Treg levels in a sustained manner. Clinically, all doses of Treg therapy tested were safe with no adverse infusion related side effects, infections or rejection events up to two years post-transplant. This study provides the necessary safety data to advance Treg cell therapy to phase II efficacy trials.


Relationships between biomechanics, tendon pathology, and function in individuals with lateral epicondylosis.
Single-cohort descriptive and correlational study. To investigate the relationships between tendon pathology, biomechanical measures, and self-reported pain and function in individuals with chronic lateral epicondylosis. Lateral epicondylosis has a multifactorial etiology and its pathophysiology is not well understood. Consequently, treatment remains challenging, and lateral epicondylosis is prone to recurrence. While tendon pathology, pain system changes, and motor impairments due to lateral epicondylosis are considered related, their relationships have not been thoroughly investigated. Twenty-six participants with either unilateral (n = 11) or bilateral (n = 15) chronic lateral epicondylosis participated in this study. Biomechanical measures (grip strength, rate of force development, and electromechanical delay) and measures of tendon pathology (magnetic resonance imaging and ultrasound) and self-reported pain and function (Patient-Rated Tennis Elbow Evaluation) were performed. Partial Spearman correlations, adjusting for covariates (age, gender, weight, and height), were used to evaluate the relationship between self-reported pain, function, and biomechanical and tendon pathology measures. Statistically significant correlations between biomechanical measures and the Patient-Rated Tennis Elbow Evaluation ranged in magnitude from 0.44 to 0.68 (P<.05); however, no significant correlation was observed between tendon pathology (magnetic resonance imaging and ultrasound) measures and the Patient-Rated Tennis Elbow Evaluation (r = -0.02 to 0.31, P>.05). Rate of force development had a stronger correlation (r = 0.54-0.68, P<.05) with self-reported function score than with grip strength (r = 0.35-0.47, P<.05) or electromechanical delay (r = 0.5, P<.05). Biomechanical measures (pain-free grip strength, rate of force development, electromechanical delay) have the potential to be used as outcome measures to monitor progress in lateral epicondylosis. In comparison, the imaging measures (magnetic resonance imaging and ultrasound) were useful for visualizing the pathophysiology of lateral epicondylosis. However, the severity of the pathophysiology was not related to pain and function, indicating that imaging measures may not provide the best clinical assessment.


An updated review of pharmacokinetic drug interactions and pharmacogenetics of statins.
Hydroxymethylglutaryl-coenzyme A reductase inhibitors (statins) lower cholesterol synthesis in patients with hypercholesterolemia. Increased statin exposure is an important risk factor for skeletal muscle toxicity. Potent inhibitors of cytochrome P450 (CYP) 3A4 significantly increase plasma concentrations of the active forms of simvastatin, lovastatin, and atorvastatin. Fluvastatin is metabolized by CYP2C9, whereas pravastatin, rosuvastatin, and pitavastatin are unaffected by inhibition by either CYP. Statins also have different affinities for membrane transporters involved in processes such as intestinal absorption, hepatic absorption, biliary excretion, and renal excretion. In this review, the pharmacokinetic aspects of drug-drug interactions with statins and genetic polymorphisms of CYPs and drug transporters involved in the pharmacokinetics of statins are discussed. Understanding the mechanisms underlying statin interactions can help minimize drug interactions and reduce the adverse side effects caused by statins. Since recent studies have shown the involvement of drug transporters such as OATP and BCRP as well as CYPs in statin pharmacokinetics, further clinical studies focusing on the drug transporters are necessary. The establishment of biomarkers based on novel mechanisms, such as the leakage of microRNAs into the peripheral blood associated with the muscle toxicity, is important for the early detection of statin side effects.


Plasmapheresis for refractory urticarial vasculitis in a patient with B-cell chronic lymphocytic leukemia.
Urticarial vasculitis is a form of cutaneous leukocytoclastic vasculitis clinically characterized by persistent and often painful urticarial lesions. Numerous systemic diseases have been associated with urticarial vasculitis, including certain hematologic disorders. This distinctive form of cutaneous necrotizing vasculitis can be resistant to standard therapeutic modalities, necessitating more aggressive intervention. We report a case of refractory urticarial vasculitis developing in association with B-cell chronic lymphocytic leukemia in a 46-year-old man. We also reviewed the literature to identify other cases of urticarial vasculitis managed with this therapeutic modality. The disease progressively improved during 6 treatments with plasmapheresis (plasma exchange). In the additional cases identified in the literature, plasmapheresis was generally effective and well tolerated. On the basis of these findings, we propose that plasmapheresis be considered a treatment option for refractory urticarial vasculitis.


Shift work and cardiovascular disease.
This paper reviews a number of studies which have presented results on the association between shift work and cardiovascular disease. It is suggested that many of the early studies suffer from methodological flaws which render them difficult to interpret. In studies in which incidence of disease has been computed and related to exposure to shift work, the results indicate a higher risk for cardiovascular disease among shift workers as compared to day workers. The evidence cannot yet, however, be considered conclusive.


Degenerative lumbar spondylolisthesis: cohort of 670 patients, and proposal of a new classification.
Degenerative spondylolisthesis is common in adults. No consensus is available about the analysis or surgical treatment of degenerative spondylolisthesis. In 2013, the French Society for Spine Surgery (Societe francaise de chirurgie du rachis) held a round table discussion to develop a classification system and assess the outcomes of the main surgical treatments. A multicentre study was conducted in nine centres located throughout France and Luxembourg. We established a database on a prospective cohort of 260 patients included between July 2011 and July 2012 and a retrospective cohort of 410 patients included in personal databases between 2009 and 2013. For patients in the prospective cohort clinical assessments were performed before and after surgery using the self-administered functional impact questionnaire AQS, SF12, and Oswestry Disability Index (ODI). Type of treatment and complications were recorded. Antero-posterior and lateral full-length radiographs were used to measure lumbar lordosis (LL), segmental lordosis (SL), pelvic incidence (PI), pelvic tilt (PT), sagittal vertical axis (SVA), and percentage of vertebral slippage. Mean follow-up was 10 months. We started a randomised clinical trial comparing posterior fusion of degenerative spondylolisthesis with versus without an inter-body cage. 60 patients were included, 30 underwent 180° fusion and 30 underwent 360° fusion using an inter-body cage implanted via a transforaminal approach. We evaluated the quality of neural decompression achieved by minimally invasive fusion technique. In a subgroup of 24 patients computed tomography (CT) was performed before and after the procedure and then compared. Mean age was 67 years and 73% of degenerative spondylolisthesis were located at L4-L5 level. The many surgical procedures performed in the prospective cohort were posterior fusion (39%), posterior fusion combined with inter-body fusion (36%), dynamic stabilization (15%), anterior lumbar fusion (8%), and postero-lateral fusion without exogenous material (2%). Peri-operative complications of any severity occurred in 17% of patients. The AQS, ODI and SF12 scores were improved significantly at follow-up. We found no differences in clinical improvements across surgical procedure types. Circumferential fusion (360°) was associated with greater relief of nerve root pain and better lordosis recovery after 1 year compared to postero-lateral fusion (180°). Post-operative CT images showed effective decompression of nervous structures after minimally invasive fusion. Longer follow-up of our patients is needed to assess the stability of the results of the various surgical procedures. Based on a radiological analysis, the authors propose a new classification with five types of degenerative spondylolisthesis: type 1, SL>5° and LL>PI-10°; type 2, SL<5° and LL>PI-10°; type 3, LL<PI-10°; type 4, LL<PI-10° and compensated sagittal balance with PT>25°; and type 5, sagittal imbalance with SVA>4 cm. IV Observational cohort study. Retrospective review of prospectively collected outcome data.


Association Between Pseudoprogression of Vestibular Schwannoma After Radiosurgery and Radiological Features of Solid and Cystic Components.
The pathophysiology of vestibular schwannoma (VS) pseudoprogression after Gamma Knife radiosurgery (GKRS) remains unclear. Radiological features in pretreatment magnetic resonance images may help predict VS pseudoprogression. This study used VS radiological features quantified using an automated segmentation algorithm to predict pseudoprogression after GKRS treatment. This is a retrospective study comprising 330 patients with VS who received GKRS. After image preprocessing and T2W/contrast-enhanced T1-weighted image (CET1W) image generation, with fuzzy C-means clustering, VSs were segmented into solid and cystic components and classified as solid and cystic. Relevant radiological features were then extracted. The response to GKRS was classified into "nonpseudoprogression" and "pseudoprogression/fluctuation". The Z test for two proportions was used to compare solid and cystic VS for the likelihood of pseudoprogression/fluctuation. Logistic regression was used to assess the correlation between clinical variables and radiological features and response to GKRS. The likelihood of pseudoprogression/fluctuation after GKRS was significantly higher for solid VS compared with cystic VS (55% vs 31%, P < .001). For the entire VS cohort, multivariable logistic regression revealed that a lower mean tumor signal intensity (SI) in T2W/CET1W images was associated with pseudoprogression/fluctuation after GKRS ( P = .001). For the solid VS subgroup, a lower mean tumor SI in T2W/CET1W images ( P = .035) was associated with pseudoprogression/fluctuation after GKRS. For the cystic VS subgroup, a lower mean SI of the cystic component in T2W/CET1W images ( P = .040) was associated with pseudoprogression/fluctuation after GKRS. Pseudoprogression is more likely to occur in solid VS compared with cystic VS. Quantitative radiological features in pretreatment magnetic resonance images were associated with pseudoprogression after GKRS. In T2W/CET1W images, solid VS with a lower mean tumor SI and cystic VS with a lower mean SI of cystic component were more likely to have pseudoprogression after GKRS. These radiological features can help predict the likelihood of pseudoprogression after GKRS.


Determining the DNA sequence elements required for binding integration host factor to two different target sites.
Binding sites for the Escherichia coli protein integration host factor (IHF) include a set of conserved bases that can be summarized by the consensus sequence WATCAANNNNTTR (W is dA or dT, R is dA or dG, and N is any nucleotide). However, additional 5'-proximal bases, whose common feature is a high dA+dT content, are also thought to be required for binding at some sites. We examine the relative contribution of these two sequence elements to IHF binding to the H' and H1 sites in attP of bacteriophage lambda by using the bacteriophage P22-based challenge-phage system. IHF was unable to act as a repressor in the challenge-phage assay at H' sites containing the core consensus element but lacking the dA+dT-rich element. This indicates that both elements are required for IHF to bind to the H' site. In contrast, the core consensus determinant alone is sufficient for IHF binding to the H1 site, which lacks an upstream dA+dT-rich region. Fifty mutants that decreased or eliminated IHF binding to the H1 site were isolated. Sequence analysis showed changes in the bases in the core consensus element only, further indicating that this determinant is sufficient for IHF binding to the H1 site. We found that placement of a dA+dT-rich element upstream of the H1 core consensus element significantly increased the affinity, suggesting that the presence of a dA+dT-rich element enhances IHF binding.


Does improved access to care affect utilization and costs for patients with chronic conditions?
To determine whether a major improvement in access (ie, implementing an open access system) in a large multispecialty medical group during 2000 was associated with changes in utilization or costs for patients with diabetes, coronary heart disease (CHD), or depression. Multilevel regression analysis of health plan administrative data. Approximately 7000 patients with diabetes, 3800 with CHD, and 6000 with depression who received all of their care in this care system served as the subjects for this study. Utilization and costs between 1999 and 2001 (before and after implementation of open access) were compared for these patients. The main outcome measures were rates of inpatient admissions and various types of outpatient encounters as well as associated costs for these subjects. Between 1999 and 2001, total office visit changes were small and varied with condition, but the proportion of these visits made to primary care physicians increased significantly by an absolute 5% to 9% and primary care physician continuity increased for each condition. Urgent care visits also decreased significantly by an absolute 5% to 9%, but there was no change in emergency department visits or hospital admissions. Total costs of care for these patients were much larger than those for the overall population of the medical group, but increased at a similar rate. A major improvement in patient access to primary care clinics was associated with increased use and continuity of primary care for patients with 3 chronic conditions, but did not affect overall resource use.


β-elemene induces caspase-dependent apoptosis in human glioma cells in vitro through the upregulation of Bax and Fas/ FasL and downregulation of Bcl-2.
β-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of β-elemene against glioma cells remains unclear. In the present study, we assessed effects of β-elemene on human glioma cells and explored the underlying mechanism. Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of β-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of β-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and β-elemne treatment group. With increase in the concentration of β-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability (IC50) was 48.5 μg/mL for 24h. β-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with β-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by β-elemene in a time and does- dependent manner. Our results indicate that β-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.


Pre-eclamptic pregnancies: an opportunity to identify women at risk for future cardiovascular disease.
Evaluation of: Bellamy L, Casas JP, Hingorani AD, Williams DJ: Pre-eclampsia and risk of cardiovascular disease and cancer in later life: systematic review and meta-analysis. Br. Med. J. 335(7627), 974 (2007). Evidence has emerged over the years suggesting that women who develop hypertensive pregnancy disorders, most notably pre-eclampsia, are at an increased risk for cardiovascular disease later in life. In this study, a systematic review and meta-analysis were performed, assessing the future risks of cardiovascular disease, cancer and all-cause mortality in women with a history of pre-eclampsia and eclampsia. Women with a history of pre-eclampsia or eclampsia, compared with women without such a history, had an increased risk for cardiovascular disease, including a fourfold increased risk for hypertension, a twofold increased risk for ischemic heart disease, stroke and deep venous thrombosis, and a 1.5-times higher all-cause mortality. The study suggests that affected women may be eligible for preventive therapies at an earlier age, especially if future studies establish the role of pre-eclampsia as an independent cardiovascular risk factor.


Safe injection practice among health care workers, Gharbiya, Egypt.
A cross-sectional study was conducted in 25 health care facilities in Gharbiya governorate to assess safe injection practices among health care workers (HCWs). Two questionnaires, one to collect information about administrative issues related to safe injection and the other to collect data about giving injections, exposure to needle stick injuries, hepatitis B vaccination status and safe injection training. Practices of injections were observed using a standardized checklist. The study revealed that there was lack of both national and local infection control policies and lack of most of the supplies needed for safe injection practices. Many safe practices were infrequent as proper needle manipulation before disposal (41%), safe needle disposal (47.5%), reuse of used syringe & needle (13.2%) and safe syringe disposal (0%). Exposure to needle stick injuries were common among the interviewed HCWs (66.2%) and hand washing was the common post exposure prophylaxis measure (63.4%). Only 11.3% of HCWs had full course hepatitis B vaccination. Infection control -including safe injections- training programs should be afforded to all HCWs.


Molecular epidemiology and emergence of Rift Valley fever.
Rift Valley fever (RVF) is a mosquito-borne viral disease which manifested itself during recent epidemics and revealed its significant potential of emergence. Studies on molecular epidemiology undertaken to better understand the factors leading to RVF emergence, have confirmed the mode of circulation of the virus and highlighted probable risks and obstacles for prevention and control. As for several other viral agents, molecular epidemiology is becoming a useful tool in the study of the emergence of RVF as a serious infectious disease.


[Five years of the National Policy of Humanization: the trajectory of a public policy].
This article presents the context of the emergence and implementation of the National Policy of Humanization (NPH), which was devised as a public policy to address and overcome the challenges perceived by Brazilian society regarding the quality and dignity of healthcare. The aims include redesigning and joint initiatives for the humanization of care, and providing solutions for problems in the field of management and organization of the health workforce. Highlighting it as a public policy, the authors analyze the main focus and theoretical and methodological approaches to cope with problems in management and practice of healthcare, signaling the need to combine strategies in both macro and micro policy areas. The analysis focuses on changes in the composition and strategies of action, which include social mobilization, support to management systems, services and health teams and the development of training processes for institutional stakeholders. The article presents results obtained in the first five years, as well as its limitations and prospects, especially the challenge facing NPH to be seen as a public policy, which cannot be achieved without the mobilization of social forces beyond the State.


Microbial life in ridge flank crustal fluids.
To determine the microbial community diversity within old oceanic crust, a novel sampling strategy was used to collect crustal fluids at Baby Bare Seamount, a 3.5 Ma old outcrop located in the north-east Pacific Ocean on the eastern flank of the Juan de Fuca Ridge. Stainless steel probes were driven directly into the igneous ocean crust to obtain samples of ridge flank crustal fluids. Genetic signatures and enrichment cultures of microorganisms demonstrate that these crustal fluids host a microbial community composed of species indigenous to the subseafloor, including anaerobic thermophiles, and species from other deep-sea habitats, such as seawater and sediments. Evidence using molecular techniques indicates the presence of a relatively small but active microbial population, dominated by bacteria. The microbial community diversity found in the crustal fluids may indicate habitat variability in old oceanic crust, with inputs of nutrients from seawater, sediment pore-water fluids and possibly hydrothermal sources. This report further supports the presence of an indigenous microbial community in ridge flank crustal fluids and advances our understanding of the potential physiological and phylogenetic diversity of this community.


Purification and in vitro antioxidant activities of tellurium-containing phycobiliproteins from tellurium-enriched Spirulina platensis.
Tellurium-containing phycocyanin (Te-PC) and allophycocyanin (Te-APC), two organic tellurium (Te) species, were purified from tellurium-enriched Spirulina platensis by a fast protein liquid chromatographic method. It was found that the incorporation of Te into the peptides enhanced the antioxidant activities of both phycobiliproteins. With fractionation by ammonium sulfate precipitation and hydroxylapatite chromatography, Te-PC and Te-APC could be effectively separated with high purity, and Te concentrations were 611.1 and 625.3 μg g(-1) protein in Te-PC and Te-APC, respectively. The subunits in the proteins were identified by using MALDI-TOF-TOF mass spectrometry. Te incorporation enhanced the antioxidant activities of both phycobiliproteins, as examined by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid assay. Moreover, Te-PC and Te-APC showed dose-dependent protection on erythrocytes against the water-soluble free radical initiator 2,2'-azo(2-asmidinopropane)dihydrochloride-induced hemolysis. In the hepatoprotective model, apoptotic cell death and nuclear condensation induced by tert-butyl hydroperoxide in HepG2 cells was significantly attenuated by Te-PC and Te-APC. Taken together, these results suggest that Te-PC and Te-APC are promising Te-containing proteins with application potential for treatment of diseases related to oxidative stress.


Screening trematodes for novel intervention targets: a proteomic and immunological comparison of Schistosoma haematobium, Schistosoma bovis and Echinostoma caproni.
With the current paucity of vaccine targets for parasitic diseases, particularly those in childhood, the aim of this study was to compare protein expression and immune cross-reactivity between the trematodes Schistosoma haematobium, S. bovis and Echinostoma caproni in the hope of identifying novel intervention targets. Native adult parasite proteins were separated by 2-dimensional gel electrophoresis and identified through electrospray ionisation tandem mass spectrometry to produce a reference gel. Proteins from differential gel electrophoresis analyses of the three parasite proteomes were compared and screened against sera from hamsters infected with S. haematobium and E. caproni following 2-dimensional Western blotting. Differential protein expression between the three species was observed with circa 5% of proteins from S. haematobium showing expression up-regulation compared to the other two species. There was 91% similarity between the proteomes of the two Schistosoma species and 81% and 78·6% similarity between S. haematobium and S. bovis versus E. caproni, respectively. Although there were some common cross-species antigens, species-species targets were revealed which, despite evolutionary homology, could be due to phenotypic plasticity arising from different host-parasite relationships. Nevertheless, this approach helps to identify novel intervention targets which could be used as broad-spectrum candidates for future use in human and veterinary vaccines.


[Inhibitory and immunological profile of therapeutic serum protein solutions].
Biseko is prepared from pooled human plasma by specific stepwise adsorption of the coagulation factors avoiding spontaneous coagulation. Biseko is manufactured from pooled plasma from more than 1000 donors. In order to ensure its hepatitis safety, the starting plasma is cold sterilized by beta-propiolactone and UV-irradiation. The inhibitory and immunological profile of the cold sterilized Biseko was compared with another commercial serum preserve and normal serum. alpha 1-antitrypsin, alpha 2-macroglobulin and antithrombin III are present in Biseko and normal serum in their biologically active forms. A certain amount of the opsonin, fibronectin, is heparin-precipitable in normal serum and has thus retained its native character, while the fibronectin in the commercial serum preserve examined is not heparin-precipitable. Biseko contains fibronectin only in trace amounts. The IgG, IgA and IgM immunoglobulin concentrations and activities in the serum preserves are equivalent to those in normal serum. One major difference between normal serum and the cold sterilized Biseko is that metabolites from the coagulation pathways are absent in Biseko. Normal serum is not suitable for therapeutic purposes because of activated enzymes formed during coagulation. The chemical analysis of the protein pattern in Biseko resembles more fresh frozen plasma without coagulation factors than normal serum.


3-Methylcholanthrene displays dual effects on estrogen receptor (ER) alpha and ER beta signaling in a cell-type specific fashion.
The biological effects of 17beta-estradiol (E(2)) are mediated by the two estrogen receptor (ER) isoforms ERalpha and ERbeta. These receptors are ligand-inducible transcription factors that belong to the nuclear receptor superfamily. These receptors are also targets for a broad range of natural and synthetic compounds that induce ER activity, including dietary compounds, pharmaceuticals, and various types of environmental pollutants such as bisphenols and polychlorinated hydroxy-biphenyls. Here, we study the effect of the combustion byproduct 3-methylcholanthrene (3-MC) on ERalpha and ERbeta. 3-MC is a compound identified previously as an activator of the aryl hydrocarbon receptor (AhR). Activation of AhR is traditionally associated with an inhibition of the E(2) signaling network. In this study, we demonstrate that 3-MC is a cell-specific activator or inhibitor of E(2) signaling pathways. We show that 3-MC acts as a repressor in some cells, presumably via the AhR, whereas it is a potent activator of ER activity in other cells. It is interesting that we demonstrate that the estrogenic effects of 3-MC are dependent on the ability of cells to metabolize parental 3-MC to alternative compounds. In summary, our results suggest that exposure to AhR ligands like 3-MC can lead to either activation or repression of E(2) signaling, depending on the cellular context.


Safety and efficacy of lamivudine in patients with severe acute or fulminant hepatitis B, a multicenter experience.
Acute hepatitis B progresses to liver failure with the need of liver transplantation in about 1% of cases. We treated patients with severe acute or fulminant hepatitis B with lamivudine in an attempt to prevent hepatitis B virus (HBV) reinfection after potential liver transplantation. Since September 2000, 17 patients with severe acute or fulminant HBV infection were treated with 100 or 150 mg lamivudine daily once we had evidence for a severe course as indicated by an INR >2.0. These were compared to a historic control from our unit and to external patients. Fourteen of the 17 patients (82.4%) survived with full recovery without liver transplantation. All these 14 individuals cleared HBsAg on lamivudine within less than 6 months. Twelve patients recovered quickly as indicated by a normalized prothrombin time within 1 week while two patients had a more prolonged course. None of the patients showed an adverse event. Three patients requiring transplantation despite lamivudine therapy had more advanced disease on admission, of whom one had additionally ingested paracetamol (acetaminophen) while the second was already HBV-DNA negative by polymerase chain reaction on admission. The lamivudine treated patients had significant higher frequency of survival without liver transplantation 82.4 vs 20% (4/20) in the historic control (P < 0.001). Similar data were derived from external centres using lamivudine (15/20, 75%). Lamivudine is safe in patients with severe acute or fulminant hepatitis B, leading to fast recovery with the potential to prevent liver failure and liver transplantation when administered early enough.


An illustrative case and a review on the dosing of recombinant factor VIIa in congenital factor XI deficiency.
Recombinant activated factor VII (rFVIIa) has been used in a very limited number of patients with severe factor XI (FXI) deficiency. The dose and duration of treatment has varied greatly between these case reports. In a few of these cases there was also evidence of thrombotic complications. We present here a report on one additional patient with congenital FXI deficiency. For two major orthopaedic procedures in this patient we used rFVIIa as a single bolus dose followed by continuous infusion at a low rate. The data from these treatment episodes, together with those from a review of the published cases, lend support to the concept of using much lower doses than in haemophilia with inhibitors. A bolus dose of 20 microg kg(-1) and thereafter maintenance of the FVII activity at approximately 3 IU mL(-1) appears effective and safe.


Fungal phytopathogens encode functional homologues of plant rapid alkalinization factor (RALF) peptides.
In this article, we describe the presence of genes encoding close homologues of an endogenous plant peptide, rapid alkalinization factor (RALF), within the genomes of 26 species of phytopathogenic fungi. Members of the RALF family are key growth factors in plants, and the sequence of the RALF active region is well conserved between plant and fungal proteins. RALF1-like sequences were observed in most cases; however, RALF27-like sequences were present in the Sphaerulina musiva and Septoria populicola genomes. These two species are pathogens of poplar and, interestingly, the closest relative to their respective RALF genes is a poplar RALF27-like sequence. RALF peptides control cellular expansion during plant development, but were originally defined on the basis of their ability to induce rapid alkalinization in tobacco cell cultures. To test whether the fungal RALF peptides were biologically active in plants, we synthesized RALF peptides corresponding to those encoded by two sequenced genomes of the tomato pathogen Fusarium oxysporum f. sp. lycopersici. One of these peptides inhibited the growth of tomato seedlings and elicited responses in tomato and Nicotiana benthamiana typical of endogenous plant RALF peptides (reactive oxygen species burst, induced alkalinization and mitogen-activated protein kinase activation). Gene expression analysis confirmed that a RALF-encoding gene in F. oxysporum f. sp. lycopersici was expressed during infection on tomato. However, a subsequent reverse genetics approach revealed that the RALF peptide was not required by F. oxysporum f. sp. lycopersici for infection on tomato roots. This study has demonstrated the presence of functionally active RALF peptides encoded within phytopathogens that harbour an as yet undetermined role in plant-pathogen interactions.


Intra-operative biopsy in chronic sinusitis detects pathogenic Escherichia coli that carry fimG/H, fyuA and agn43 genes coding biofilm formation.
The aim of this study was to investigate whether or not surgical biopsy of sinus tissue in chronic sinusitis, not responsive to treatment, would detect E. coli. We intended to evaluate E. coli virulence genes, therefore dispute the causal role of such an unusual microorganism in chronic sinusitis, as well as consider effective pathogen-targeted therapy. Patients with E. coli isolated by intra-operative puncture biopsy were included in the study. Genetic analysis of E. coli isolates, including phylogenetic grouping and virulence factor characteristics, were done by multiplex PCR. We identified 26 patients with chronic sinusitis, in which 26 E. coli isolates were cultured. The E. coli isolates belonged mainly to pathogenic phylogenetic group B2, and carried multiple virulence genes. Three genes in particular were present in all (100%) of examined isolates, they were (1) marker agn43 gene for forming biofilm, (2) type 1 fimbriae (fimG/H gene) and (3) yersiniabactin receptor (fyuA). Furthermore, a pseudo-phylogenetic tree of virulence genes distribution revealed possible cooperation between agn43, fimG/H, and fyuA in the coding of biofilm formation. Intra-operative-biopsy and culture-based therapy, targeting the isolated E. coli, coincided with long-term resolution of symptoms. This is the first report demonstrating an association between a highly pathogenic E. coli, chronic sinus infection, and resolution of symptoms upon E. coli targeted therapy, a significant finding due to the fact that E. coli has not been considered to be a commensal organism of the oropharynx or sinuses. We postulate that the simultaneous presence of three genes, each coding biofilm formation, may in part account for the chronicity of E. coli sinusitis.


Physical activity and inactivity in relation to sex hormone, prolactin, and insulin-like growth factor concentrations in premenopausal women - exercise and premenopausal hormones.
An association between physical activity and premenopausal breast cancer risk may be due, in part, to relationships with sex hormones or growth factors. Therefore, we assessed whether MET-h/week of total physical activity (moderate-to-vigorous intensity), walking, or vigorous physical activity, and h/week of standing or sitting were associated with plasma concentrations of several hormones. We examined levels of estrogens, androgens, progesterone, prolactin, sex hormone binding globulin (SHBG), insulin-like growth factor-1 (IGF-1), IGF binding protein-3, and growth hormone (GH) in 565 premenopausal women, ages 33-52 years, from the Nurses' Health Study II (NHSII). About 87% of women had both timed follicular and luteal samples; other women had one untimed sample. In general we observed few associations between sex hormone or IGF levels and measures of physical activity or inactivity. However, free testosterone was modestly inversely associated with total physical activity (p-trend = 0.02). Luteal estradiol, free estradiol, and estrone also were inversely associated with total physical activity (p-trend = 0.10, 0.04, 0.01, respectively); however, the trend was substantially attenuated when excluding women with anovulatory cycles or irregular cycles. These cross-sectional results suggest that physical activity and inactivity have limited associations with premenopausal sex hormone and growth factor levels, except possibly luteal estrogens.


Measurement of liver blood flow by galactose clearance.
Low-dose galactose clearance is a new method for measuring functional (nutrient) liver blood flow. In 22 healthy beagle dogs, the mean (+/- SD) blood galactose clearance rate of 311 +/- 93 mL/min was not significantly different from the mean measurement obtained using electromagnetic flow probes (322 +/- 37 mL/min). This shows that galactose clearance can be used to measure liver blood flow in healthy dogs. The 22 dogs were divided into two groups of 11. The first group underwent portacaval shunting and weekly galactose clearance rates were measured until death an average of 6 weeks later. The anticipated fall in liver blood flow was successfully detected by the second week after shunting. This suggests that long-term (week-to-week) changes in liver blood flow can be detected by this method. In the second group, ligation of the common bile duct was used to induce secondary biliary cirrhosis. Galactose clearance was measured weekly for 6 weeks and showed a significant decrease by 6 weeks. At 7 weeks, laparotomy was performed in order to take flow-probe measurements; the galactose clearance rate was also measured. Whereas the two methods were similar at the time of the original operation, 7 weeks after ligation there was a significant difference (p = 0.02) with the rate of liver blood flow as measured by galactose clearance being much lower than the flow rate measured by the electromagnetic flow probes. These findings suggest that in cirrhotic dogs, galactose clearance measures functional or effective (nutrient) liver blood flow whereas the electromagnetic flow probe measures anatomic flow. The effective flow gives a more accurate reflection of perfusion of the hepatocyte by blood.


Increased Functional Connectivity in an Insula-Based Network is Associated with Improved Smoking Cessation Outcomes.
Little is known regarding the underlying neurobiology of smoking cessation. Neuroimaging studies indicate a role for the insula in connecting the interoceptive awareness of tobacco craving with a larger brain network that motivates smoking. We investigated differences in insula-based functional connectivity between smokers who did not relapse during a quit attempt vs those who relapsed. Smokers (n=85) underwent a resting-state functional connectivity scan and were then randomized into two groups (either smoking usual brand cigarettes or smoking very low nicotine cigarettes plus nicotine replacement therapy) for 30 days before their target quit date. Following the quit date, all participants received nicotine replacement therapy and their smoking behavior was observed for 10 weeks. Participants were subsequently classified as nonrelapsed (n=44) or relapsed (i.e., seven consecutive days of smoking ⩾1 cigarette/day; n=41). The right and left insula, as well as insula subdivisions (posterior, ventroanterior, and dorsoanterior) were used as seed regions of interest in the connectivity analysis. Using the right and left whole-insula seed regions, the nonrelapsed group had greater functional connectivity than the relapsed group with the bilateral pre- and postcentral gyri. This effect was isolated to the right and left posterior insula seed regions. Our results suggest that relapse vulnerability is associated with weaker connectivity between the posterior insula and primary sensorimotor cortices. Perhaps greater connectivity in this network improves the ability to inhibit a motor response to cigarette cravings when those cravings conflict with a goal to remain abstinent. These results are consistent with recent studies demonstrating a positive relationship between insula-related functional connectivity and cessation likelihood among neurologically intact smokers.


Self-Attractive Hartree Decomposition: Partitioning Electron Density into Smooth Localized Fragments.
Chemical bonding plays a central role in the description and understanding of chemistry. Many methods have been proposed to extract information about bonding from quantum chemical calculations, the majority of them resorting to molecular orbitals as basic descriptors. Here, we present a method called self-attractive Hartree (SAH) decomposition to unravel pairs of electrons directly from the electron density, which unlike molecular orbitals is a well-defined observable that can be accessed experimentally. The key idea is to partition the density into a sum of one-electron fragments that simultaneously maximize the self-repulsion and maintain regular shapes. This leads to a set of rather unusual equations in which every electron experiences self-attractive Hartree potential in addition to an external potential common for all the electrons. The resulting symmetry breaking and localization are surprisingly consistent with chemical intuition. SAH decomposition is also shown to be effective in visualization of single/multiple bonds, lone pairs, and unusual bonds due to the smooth nature of fragment densities. Furthermore, we demonstrate that it can be used to identify specific chemical bonds in molecular complexes and provides a simple and accurate electrostatic model of hydrogen bonding.


Detailed Long-Term Follow-Up of Patients Who Relapsed After the Nordic Mantle Cell Lymphoma Trials: MCL2 and MCL3.
Mantle cell lymphoma (MCL) is an incurable disease with a highly variable clinical course. The prognosis after relapse is generally poor, and no standard of care exists. We investigated the postrelapse outcomes of 149 patients who were initially treated in the Nordic Lymphoma Group trials, MCL2 or MCL3, both representing intensive cytarabine-containing frontline regimens including autologous stem cell transplant. Patients with progression of disease before 24 months (POD24, n = 51, 34%) displayed a median overall survival of 6.6 months compared with 46 months for patients with later POD (n = 98, 66%; P < 0.001). MCL international prognostic index, cell proliferation marker, blastoid morphology, and TP53 mutations showed independent prognostic value irrespective of POD24, and in a combined, exploratory risk score, patients with 0, 1, 2-3, or 4-5 high-risk markers, respectively, displayed a 5-year overall survival of 62%, 39%, 31%, and 0%. By a comparison of median progression-free survival of the different salvage therapies in the relapse setting, bendamustine-rituximab was superior to all other combination chemotherapy regimens; however, it was also associated with longer responses to last line of therapy. Collectively, we confirm the prognostic impact of POD24 and highlight the relevance of other biomarkers, and we emphasize the importance of novel therapies for patients with high-risk features at first POD.


From filters to features: scale-space analysis of edge and blur coding in human vision.
To make vision possible, the visual nervous system must represent the most informative features in the light pattern captured by the eye. Here we use Gaussian scale-space theory to derive a multiscale model for edge analysis and we test it in perceptual experiments. At all scales there are two stages of spatial filtering. An odd-symmetric, Gaussian first derivative filter provides the input to a Gaussian second derivative filter. Crucially, the output at each stage is half-wave rectified before feeding forward to the next. This creates nonlinear channels selectively responsive to one edge polarity while suppressing spurious or "phantom" edges. The two stages have properties analogous to simple and complex cells in the visual cortex. Edges are found as peaks in a scale-space response map that is the output of the second stage. The position and scale of the peak response identify the location and blur of the edge. The model predicts remarkably accurately our results on human perception of edge location and blur for a wide range of luminance profiles, including the surprising finding that blurred edges look sharper when their length is made shorter. The model enhances our understanding of early vision by integrating computational, physiological, and psychophysical approaches.


Perceptions of patients with wounds due to chronic limb-threatening ischemia.
Patients with chronic limb-threatening ischemia (CLTI) face numerous barriers to caring for lower extremity wounds. We explored the perceptions of CLTI patients to their wound/management and sought to determine attitudes towards their vascular provider as well as willingness for management through telemedicine. Patients admitted to hospital for treatment of Rutherford Grade 5 and 6 CLTI were asked complete a wound evaluation survey and took part in a semi-structured interview. Semi-structured interviews were recorded, transcribed, and analyzed using an inductive coding strategy. Codes were grouped for thematic analysis and aggregated into assertions. Eleven patients with a mean age of 60 years (35-79 years) were interviewed. All patients had peripheral artery disease (PAD) and eight patients had diabetes as well. Three overarching themes were identified. First, patients appear to have limited coping mechanisms and are overwhelmed by the care of their wounds. Second, in this cohort of patients, many had become passive observers of their care as demonstrated by a limited understanding of their disease processes and detachment from wound management. The third theme was how strong the desire to do everything to prevent limb loss was, but patients acknowledged this is hard to translate into real life with limited resources. Patients with CLTI have concerns that vascular providers must recognize and address to build strong patient-provider relationships and increase activation for management of their wounds and other medical conditions. Patients who have access to technology and with guidance may be able to understand getting care through remote medicine.


Feasibility in the echocardiographic estimation of parameters of the right ventricle in prone position.
Echocardiographic cardiac parameters in the prone position are usually obtained with an esophageal probe. The feasibility of obtaining them by means of a transthoracic approach is unknown. Estimating the feasibility to obtain parameters of the right ventricle by transthoracic echocardiography in prone position on the subject. Pilot design of consecutive case series without cardiopulmonary disease. Demographic, vital signs and echocardiographic variables were defined in the ventral initial, prone and ventral final decubitus positions. The data are shown with averages and standard deviations, and frequencies and percentages according to the variable. The differences between the positions were calculated with ANOVA of repeated samples and adjustment of Bonferroni test. Intra-subject variability was obtained by the Bland-Altman procedure and its 95% confidence interval. We studied 50 subjects, 44 (88%) males, age 30 ± 6 years and body mass index 25.65 ± 2.71 kg/m2. Tricuspid annular plane systolic excursion (TAPSE) and S'-wave were measured 100% of the time. The vital signs and echocardiographic variables according to the position had differences in: heart rate (74 ± 9 vs. 77 ± 9 vs. 75 ± 8 beats/min), partial oxygen saturation (94.40 ± 1.70 vs. 96.64 ± 1.79 vs. 95.32 ± 1.36%) and mean systemic blood pressure (65.33 ± 5.38 vs. 67.69 ± 6.31 vs. 65.29 ± 5.62 mmHg); TAPSE (19.74 ± 3.24 vs. 21.60 ± 2.97 vs. 19.44 ± 2.84 mm), mean difference (bias) 0 (2, -2.0) and S'-wave (13.52 ± 1.87 vs. 15.02 ± 2.09 vs. 13.46 ± 1.55 cm/s), mean difference (bias) -0.46 (1.21, -2.14) respectively. Obtaining right ventricle parameters by transthoracic ecocardiopraphy is feasible in the prone position.


Cold Bonding Method for Metallic Powder Coatings.
An efficient and simple method for preparing bonded metallic powder coating is in high demand in the paint manufacturing and application industries. The bonding purpose is to keep the mass percentage of metallic pigment consistent between the original and recycled coating powder, which aims at solving the problem of recyclability. One possible method capable of realizing this goal is using the binder to cohere metallic pigment with base particles through a cold bonding method. Through this approach, the pre-curing and high-reject-rate problems generally present in thermal bonding can be completely eliminated. In this paper, polyacrylic acid (PAA) and polyvinyl alcohol (PVA) are applied as binders for the bonding process. At various dosages of liquid binder and D.I. water, bonded samples with different bonding effect were prepared. Finally, a good bonding quality with the lowest variance between the mass concentrations of Al flakes in the original powder (before spray) and deposited powder (after spray) 2.94% with PAA as a binder and 0.46% with PVA as a binder was achieved. These results manifest that the cold bonding method is a green and simple approach for preparing the metallic powder coating.


Off-policy reinforcement learning for H∞ control design.
The H∞ control design problem is considered for nonlinear systems with unknown internal system model. It is known that the nonlinear H∞ control problem can be transformed into solving the so-called Hamilton-Jacobi-Isaacs (HJI) equation, which is a nonlinear partial differential equation that is generally impossible to be solved analytically. Even worse, model-based approaches cannot be used for approximately solving HJI equation, when the accurate system model is unavailable or costly to obtain in practice. To overcome these difficulties, an off-policy reinforcement leaning (RL) method is introduced to learn the solution of HJI equation from real system data instead of mathematical system model, and its convergence is proved. In the off-policy RL method, the system data can be generated with arbitrary policies rather than the evaluating policy, which is extremely important and promising for practical systems. For implementation purpose, a neural network (NN)-based actor-critic structure is employed and a least-square NN weight update algorithm is derived based on the method of weighted residuals. Finally, the developed NN-based off-policy RL method is tested on a linear F16 aircraft plant, and further applied to a rotational/translational actuator system.


Changes in the Electrophysical Parameters of Nanomodified Elastomers Caused by Electric Current's Passage.
The development of reliable and effective functional materials that can be used in various technological fields and environmental conditions is one of the goals of modern nanotechnology. Heating elements' manufacturing requires understanding the laws of heat transfer under conditions of different supply voltages, as this expands the possibilities of such materials' application. Elastomers based on silicon-organic compounds and polyurethane modified with multi-walled carbon nanotubes (MWCNTs) were studied at various concentrations of Ni/MgO or Co-Mo/MgO and voltages (220, 250, and 300 V). It was found that an increase in voltage from 220 to 300 V leads to an initial increase in specific power on one-third followed by a subsequent decrease in a specific power when switched on again to 220 V (for -40 °C) of up to ~44%. In turn, for a polyurethane matrix, an increase in voltage to 300 V leads to an initial peak power value of ~15% and a decrease in power when switched on again by 220 V (for -40 °C) to ~36% (Ni/MgO -MWCNT). The conducted studies have shown that the use of a polyurethane matrix reduces power degradation (associated with voltage surges above 220 V) by 2.59% for Ni/MgO-based MWCNT and by 10.42% for Co-Mo/MgO. This is due to the better heat resistance of polyurethane and the structural features of the polymer and the MWCNT. The current studies allow us to take the next step in the development of functional materials for electric heating and demonstrate the safety of using heaters at a higher voltage of up to 300 V, which does not lead to their ignition, but only causes changes in electrophysical parameters.


The MIG Chemokine in Inflammatory Myopathies.
The chemokine monokine induced by interferon (IFN)-γ (MIG) is expressed in idiopathic inflammatory myopathies muscle. Abundant expression of MIG was observed on macrophages and T cells surrounding and invading non-necrotic muscle fibers in polymyositis and in inclusion-body myositis and in T cells in perimysial infiltrates of dermatomyositis. MIG is also localized to blood vessel endothelial cells in all inflammatory and normal muscle tissues and it exerts its biological effects mainly via binding to the chemokine (C-X-C motif) receptor (CXCR)3. Serum MIG is high in patients with inflammatory myopathies. Human skeletal muscle cells might actively self-promote muscular inflammation by eliciting MIG secretion, under the influence of cytokines (IFN-γ, tumor necrosis factor-α), which can amplify Th1 cell tissue infiltration in vivo. It has been shown that drugs able to block the MIG/CXCR3 axis can suppress inflammation in muscle.


Protein synthesis inhibitors and the chemical chaperone TMAO reverse endoplasmic reticulum perturbation induced by overexpression of the iodide transporter pendrin.
An outcome of overloading of the endoplasmic reticulum (ER) folding machinery is a perturbation in ER function and the formation of intracellular aggregates. The latter is a key pathogenic factor in numerous diseases known as ER storage diseases. Here, we report that heterologous overexpression of the green fluorescent protein-tagged iodide transporter pendrin (GFP-PDS) perturbs folding and degradation processes in the ER. Pendrin (PDS) is a chloride-iodide transporter found in thyroid cells. Mutations in PDS can cause its retention in the ER and are associated with Pendred syndrome. Biochemical and live-cell analyses demonstrated that wild-type GFP-PDS is predominantly retained in perinuclear aggregates and in ER membranes, causing their collapse and vesiculation. Inhibition of protein synthesis by cycloheximide (CHX) or puromycin caused dissociation of the GFP-PDS aggregates and returned the ER to its normal reticular morphology. Blocking protein synthesis promoted folding and export of ER-retained GFP-PDS, as demonstrated by surface-biotinylation analysis and by CHX- or puromycin-induced accumulation of YFP-PDS in the Golgi apparatus during a 20 degrees C temperature-block experiment. The chemical chaperone trimethylamine-N-oxide (TMAO) also reversed the GFP-PDS-mediated ER collapse and vesiculation, suggesting that exposed hydrophobic stretches of misfolded or aggregated GFP-PDS may contribute to ER retention. These data suggest that GFP-PDS is a slow-folding protein with a propensity to form aggregates when overexpressed. Thus, we describe a system for the reversible induction of ER stress that is based entirely on the heterologous overexpression of GFP-PDS.


Association between flat-panel computed tomography hyperattenuation and clinical outcome after successful recanalization by endovascular treatment.
Hyperattenuation on CT scanning performed immediately after endovascular treatment (EVT) is known to be associated with the final infarct. As flat-panel CT (FPCT) scanning is readily accessible within their angiography suite, the authors evaluated the ability of the extent of hyperattenuation on FPCT to predict clinical outcomes after EVT. Patients with successful recanalization (modified Thrombolysis in Cerebral Infarction grade 2b or 3) were reviewed retrospectively. The extent of hyperattenuation was assessed by the Alberta Stroke Program Early CT Score on FPCT (FPCT-ASPECTS). FPCT-ASPECTS findings were compared according to functional outcome and malignant infarction. The predictive power of the FPCT-ASPECTS with initial CT images before EVT (CT-ASPECTS) and follow-up diffusion-weighted images (MR-ASPECTS) was also compared. A total of 235 patients were included. All patients were treated with mechanical thrombectomy, and 45.5% of the patients received intravenous tissue plasminogen activator. The mean (± SD) time from stroke onset to recanalization was 383 ± 290 minutes. The FPCT-ASPECTS was significantly different between patients with a favorable outcome and those without (mean 9.3 ± 0.9 vs 6.7 ± 2.6) and between patients with malignant infarction and those without (3.4 ± 2.9 vs 8.8 ± 1.4). The FPCT-ASPECTS was an independent factor for a favorable outcome (adjusted OR 3.28, 95% CI 2.12-5.01) and malignant infarction (adjusted OR 0.42, 95% CI 0.31-0.57). The area under the curve (AUC) of the FPCT-ASPECTS for a favorable outcome (0.862, cutoff ≥ 8) was significantly greater than that of the CT-ASPECTS (0.637) (p < 0.001) and comparable to that of the MR-ASPECTS (0.853) (p = 0.983). For malignant infarction, the FPCT-ASPECTS was also more predictive than the CT-ASPECTS (AUC 0.906 vs 0.552; p = 0.001) with a cutoff of ≤ 5. The FPCT-ASPECTS was highly predictive of clinical outcomes in patients with successful recanalization. FPCT could be a practical method to immediately predict clinical outcomes and thereby aid in acute management after EVT.


Piezoelectric Transformer-Based High-Voltage Pulse Generator Using Wide-Bandgap Semiconductors for Medical Electroporation Therapy.
In this paper, a new application of Piezoelectric Transformer (PT)-based power converters to generate high-voltage (HV) bipolar pulses for medical electroporation therapy is proposed. In particular, PT-based power conversion is investigated as an alternative to magnetics-based approaches of generating HV from a relatively low-voltage (LV) input source for application in electroporation therapy. The detailed PT-based system design and selection of wide bandgap semiconductor switches such as GaN FETs, high-voltage SiC diodes and SiC MOSFETs, as well as simulation results to demonstrate proof-of-concept using LTSpice are presented. Preliminary experimental results of the PT-based capacitor charger are shown, and work is ongoing to develop a complete hardware prototype of the proposed HV pulse generator.


Acute effects of ambient inhalable particles in asthmatic and nonasthmatic children.
Although increases in inhalable particle (PM10) concentrations have been associated with acute reductions in the level of lung function and increased symptom reporting in children, including children with asthma, it is not clear whether these effects occur largely in asthmatic children, or even whether asthmatic children are more likely to experience these effects than children without asthma. To address these points, the following subgroups of children were selected from a survey population of all 2,200 elementary school children (6 to 13 yr of age) in a pulp mill community on the west coast of Vancouver Island: (1) all children with physician-diagnosed asthma (n = 75 participated), (2) all children with an exercise-induced fall in FEV1 without diagnosed asthma (n = 57), (3) all children with airway obstruction (FEV1/FVC < 0.76) without either of the above (n = 18), and (4) control children without any of the above (n = 56). The children were followed for as long as 18 mo with twice daily measurements of peak expiratory flow (PEF) and daily symptom diary recording. Maximum daily PM10 concentration was 159 microm/m3 (median, 22.1), but only 8 d (1.2%) had concentrations above 100 microg/m3. In an analysis that accounted for time-varying covariates, and serially correlated and missing data, for the entire sample of children, increases in PM10 were associated with reductions in PEF and increased reporting of cough, phlegm production, and sore throat. For the subgroup of children with diagnosed asthma, PEF in the time period with the highest PM10 concentrations fell by an estimated 0.55 L/min (95% CI, 0.06 to 1.05) for a 10 microg/m3 PM10 increase above the mean daily PM10 concentration of 27.3 microg/m3 and the odds of reported cough increased by 8% (95% CI, 0 to 16%); no consistent effects were observed in the other groups of children. It is concluded that children experience reductions in PEF and increased symptoms after increases in relatively low ambient PM10 concentrations, and that children with diagnosed asthma are more susceptible to these effects than are other children.


Geographic Distribution of Mental Health Problems Among Chinese College Students During the COVID-19 Pandemic: Nationwide, Web-Based Survey Study.
Since the COVID-19 outbreak was first reported, considerable attention has been drawn to mental health problems among college students. We aimed to estimate the prevalence of anxiety and depressive symptoms among college students in different geographical areas of China during the early stage of the COVID-19 outbreak. A nationwide cross-sectional survey was conducted among Chinese college students of 16 provinces or municipalities from February 4 to 12, 2020. A web-based survey was adopted to collect information from these college students, including demographics, perceived risk of infection, attitudes toward the epidemic and its control, and mental health status. Anxiety symptoms were assessed using the Generalized Anxiety Disorder scale, and depressive symptoms were assessed using the Patient Health Questionnaire. Chi-square test was used to compare the percentage of perceived risk of infection and attitude toward COVID-19 among college students in different geographic locations. Binary logistic models were used to identify associations between geographic locations and mental health problems after controlling for covariates. A total of 11,787 participants were analyzed in this study (response rate: 79.7%). The prevalence of anxiety and depressive symptoms among college students was 17.8% (95% CI 17.1%-18.5%) and 25.9% (95% CI 25.1%-26.7%), respectively. After controlling for covariates, current residence area in Wuhan city was found to have a positive association with anxiety symptoms (odds ratio [OR] 1.37, 95% CI 1.11-1.68) and depressive symptoms (OR 1.32, 95% CI 1.09-1.59). Similarly, college location in Wuhan city was found to have a positive association with anxiety symptoms (OR 1.20, 95% CI 1.07-1.35) and depressive symptoms (OR 1.22, 95% CI 1.10-1.36). History of residence in or travel to Wuhan city in the past month was also positively associated with anxiety symptoms (OR 1.62, 95% CI 1.46-1.80) and depressive symptoms (OR 1.48, 95% CI 1.35-1.63). Furthermore, the perceived risk of COVID-19 was higher among students whose college location and current residence area were in Wuhan city, and it was positively associated with anxiety and depressive symptoms. During the COVID-19 pandemic, mental health problems among Chinese college students were widespread and geographically diverse. Our study results provide further insight for policymakers to develop targeted intervention strategies.


Conformational heterogeneity of the BTK PHTH domain drives multiple regulatory states.
Full-length BTK has been refractory to structural analysis. The nearest full-length structure of BTK to date consists of the autoinhibited SH3-SH2-kinase core. Precisely how the BTK N-terminal domains (the Pleckstrin homology/Tec homology (PHTH) domain and proline-rich regions (PRR) contain linker) contribute to BTK regulation remains unclear. We have produced crystals of full-length BTK for the first time but despite efforts to stabilize the autoinhibited state, the diffraction data still reveals only the SH3-SH2-kinase core with no electron density visible for the PHTH-PRR segment. CryoEM data of full-length BTK, on the other hand, provide the first view of the PHTH domain within full-length BTK. CryoEM reconstructions support conformational heterogeneity in the PHTH-PRR region wherein the globular PHTH domain adopts a range of states arrayed around the autoinhibited SH3-SH2-kinase core. On the way to activation, disassembly of the SH3-SH2- kinase core opens a new autoinhibitory site on the kinase domain for PHTH domain binding that is ultimately released upon interaction of PHTH with PIP3. Membrane-induced dimerization activates BTK and we present here a crystal structure of an activation loop swapped BTK kinase domain dimer that likely represents the conformational state leading to trans-autophosphorylation. Together, these data provide the first structural elucidation of full-length BTK and allow a deeper understanding of allosteric control over the BTK kinase domain during distinct stages of activation.


[Accumulation of anomalous glycopolymers in the walls of main lymphatic vessels of the extremities in patients with chronic lymphedema].
Redistribution of lectin receptor sites in the mam-lymphatics of human extremities in primary and secondary lymphedema in comparison with normal lymphatics has been investigated using conjugates to peroxidase of LCA, PNA, RCA, SBA, WGA and LAA. Chronic lymphedema caused no essential differences in lectin binding to tissue structures of intima and adventitia: endotheliocytes were strong reactive both with LCA, PNA, WGA and LAA; all lectins used bound fibrillary structures in subendothelial layer and adventitia. On the opposite, tunica media of main lymphatics in primary lymph stasis displayed accumulation of LCA-, PNA-, SBA- and LAA-binding deposits round smooth myocytes. When PNA used, the appearance of a specific "wide loop shaped reticulum" within lymphatics media in lymph stasis was observed. We consider, that lectin receptor sites accumulation phenomenon reflects some changes in smooth myocytes metabolism, possibly caused by dystrophia of these cell population with consequent incomplete glycosylation of synthesized glycopolymers.


Glyceraldehyde-3-phosphate dehydrogenase acts as a mitochondrial trans-S-nitrosylase in the heart.
Mitochondrial proteins have been shown to be common targets of S-nitrosylation (SNO), but the existence of a mitochondrial source of nitric oxide remains controversial. SNO is a nitric oxide-dependent thiol modification that can regulate protein function. Interestingly, trans-S-nitrosylation represents a potential pathway for the import of SNO into the mitochondria. The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which has been shown to act as a nuclear trans-S-nitrosylase, has also been shown to enter mitochondria. However, the function of GAPDH in the mitochondria remains unknown. Therefore, we propose the hypothesis that S-nitrosylated GAPDH (SNO-GAPDH) interacts with mitochondrial proteins as a trans-S-nitrosylase. In accordance with this hypothesis, SNO-GAPDH should be detected in mitochondrial fractions, interact with mitochondrial proteins, and increase mitochondrial SNO levels. Our results demonstrate a four-fold increase in GAPDH levels in the mitochondrial fraction of mouse hearts subjected to ischemic preconditioning, which increases SNO-GAPDH levels. Co-immunoprecipitation studies performed in mouse hearts perfused with the S-nitrosylating agent S-nitrosoglutathione (GSNO), suggest that SNO promotes the interaction of GAPDH with mitochondrial protein targets. The addition of purified SNO-GAPDH to isolated mouse heart mitochondria demonstrated the ability of SNO-GAPDH to enter the mitochondrial matrix, and to increase SNO for many mitochondrial proteins. Further, the overexpression of GAPDH in HepG2 cells increased SNO for a number of different mitochondrial proteins, including heat shock protein 60, voltage-dependent anion channel 1, and acetyl-CoA acetyltransferase, thus supporting the role of GAPDH as a potential mitochondrial trans-S-nitrosylase. In further support of this hypothesis, many of the mitochondrial SNO proteins identified with GAPDH overexpression were no longer detected with GAPDH knock-down or mutation. Therefore, our results suggest that SNO-GAPDH can act as a mitochondrial trans-S-nitrosylase, thereby conferring the transfer of SNO from the cytosol to the mitochondria.


Biofuel production improvement with genome-scale models: The role of cell composition.
Genome-scale models have developed into a vital tool for rational metabolic engineering. These models balance cofactors and energetic requirements and determine biosynthetic precursor availability in response to environmental and genetic perturbations. In particular, allocation of additional reducing power is an important strategy for engineering potential biofuel production from microbes. Many potential biofuel solvents induce biomolecular changes on the host organism that are not yet captured by genome-scale models. Here, methods of construction for several biomass constituting equations are reviewed along with potential changes to cellular composition with potential biofuels exposure. The biomass constituting equations of potential host organisms with existing genome-scale models are compared side-by-side to explore their evolution over the years and to explore differences that arise when these equations are compiled by different research groups. Genome-scale model simulation results attempt to address and provide guidance for further research into: (i) whether inconsistencies in the biomass constituting equations are relevant to predictions of solvent production, (ii) what level of detail is necessary to accurately describe cellular composition, and (iii) future developments that may enable more accurate characterizations of biomolecular composition.


The ocularists' management of congenital microphthalmos and anophthalmos.
Early socket stimulation is crucial for management of congenital anophthalmos and microphthalmos among infants. Progressive sized hard conformers and lid expansion devices can expand the small socket in these patients. The ocularists' management of these two conditions is discussed and techniques are introduced.


Remote AI Supported E-Multidisciplinary Oncology Conference in Breast Cancer as a Technology and Method to Optimize Outcomes in the Peripheries.
Feasibility-reliability control of Telemedicine Systems (TS) integrated with Multimedia Systems (MS) and Artificial intelligence (AI) for remote e-Multidisciplinary Oncology Conference in Breast Cancer. Forty (n1=40) patients suffering from breast surgical oncology malignant (n2=32) and non-malignant (n3=8) diseases classified to seven categories: Nipple Discharge, Dominant Breast Mass, Occult Breast Lesion, Early Breast Carcinoma, Advanced Breast Carcinoma, Recurrent Breast Carcinoma) and treated clinically with the standard diagnostic (Mammography, US, MRI, Cytology, Pathology, BRCA1/2 Mutation Predisposition and Breast Cancer Risk Analysis) surgical, auxiliary therapeutic methods. Then clinical decisions compared to those proposed remotely by the virtual AI supported e-Oncology Conference for each patient. In four (n4=4) out of forty patients (TS, MS and AI) supported decision making and surgical treatment proposal including postoperative Radiotherapy proposal was not as clear as expected. Non-output answer for non-malignant breast pathologies (n3=8) was accurately indicated by (MS and AI). Mean accuracy of (TS, MS and AI) for: 1.Surgical Operative Planning including Rad=94.1%, 2.Chem=96.8%, 3.Horm=96.7% [In 95%, (Confidence interval: 85-99%)]. High feasibility-reliability of the virtual AI supported e-Multidisciplinary Oncology Conference for remote decision making and surgical planning and for optimum outcomes in Breast Cancer treatment makes it a clinical necessity especially for the periphery of Hellas.


Inducible Genetic Code Expansion in Eukaryotes.
Genetic code expansion (GCE) is a versatile tool to site-specifically incorporate a noncanonical amino acid (ncAA) into a protein, for example, to perform fluorescent labeling inside living cells. To this end, an orthogonal aminoacyl-tRNA-synthetase/tRNA (RS/tRNA) pair is used to insert the ncAA in response to an amber stop codon in the protein of interest. One of the drawbacks of this system is that, in order to achieve maximum efficiency, high levels of the orthogonal tRNA are required, and this could interfere with host cell functionality. To minimize the adverse effects on the host, we have developed an inducible GCE system that enables us to switch on tRNA or RS expression when needed. In particular, we tested different promotors in the context of the T-REx or Tet-On systems to control expression of the desired orthogonal tRNA and/or RS. We discuss our result with respect to the control of GCE components as well as efficiency. We found that only the T-REx system enables simultaneous control of tRNA and RS expression.


Intra-arterial versus negative-staining of embryonal resection borders with indocyanine green (ICG) fluorescence for total mesorectal excision in colorectal cancer - an experimental feasibility study in a porcine model.
Colorectal cancer (CRC) is one of the most common malignancies worldwide. Laparoscopic lower rectal resections in accordance with the oncological principles are recommended as the gold standard for CRC surgical management. However, the learning curve for adopting these techniques is quite steep and the incomplete resections are predictive of local recurrence. This study was conducted in an attempt to find a way to help surgeons to overcome some of these difficulties and define the right resection margins. As such, we carried out two laparoscopic lower rectal resections in porcine models. The first resection was performed following the ligation and selective infusion of Indocyanine Green (ICG) into the inferior mesenteric artery (IMA), and the second after the ligation of both inferior mesenteric artery and vein (IMV) and systemic intravenous infusion of ICG. Fluorescence was detected in real time by means of an infrared imaging system. Sharp resection margins were defined after intra-arterial infusion, and all the tissues in the IMA basin were colored in the first case. In the second model every organ and tissue was colored except the rectum, urinary bladder and ductus deferens. Although systemic intra-venous application of ICG and negative-staining of the rectum including the mesorectum is much easier compared to laparoscopic inter-arterial perfusion through IMA, image results of selevtive IMA-perfusion appear in sharper discrimination of the several layers. Further investigation should focus on simplifying this technique.


Deciphering the Pharmacological Mechanism of the Herb Radix Ophiopogonis in the Treatment of Nasopharyngeal Carcinoma by Integrating iTRAQ-Coupled 2-D LC-MS/MS Analysis and Network Investigation.
The herb Radix Ophiopogonis (RO) has been used effectively to treat nasopharyngeal carcinoma (NPC) as an adjunctive therapy. Due to the complexity of the traditional Chinese herbs, the pharmacological mechanism of RO's action on NPC remains unclear. To address this problem, an integrative approach bridging proteome experiments with bioinformatics prediction was employed. First, differentially expressed protein profile from NPC serum samples was established using isobaric tag for relative and absolute quantification (iTRAQ) coupled 2-D liquid chromatography (LC)-MS/MS analysis. Second, the RO putative targets were predicted using Traditional Chinese Medicines Integrated Database and known therapeutic targets of NPC were collected from Drugbank and OMIM databases. Then, a network between RO putative targets and NPC known therapeutic targets was constructed. Third, based on pathways enrichment analysis, an integrative network was constructed using DAVID and STRING database in order to identify potential candidate targets of RO against NPC. As a result, we identified 13 differentially expressed proteins from clinical experiments compared with the healthy control. And by bioinformatics investigation, 12 putative targets of RO were selected. Upon interactions between experimental and predicted candidate targets, we identified three key candidate targets of RO against NPC: VEGFA, TP53, and HSPA8, by calculating the nodes' topological features. In conclusion, this integrative pharmacology-based analysis revealed the anti-NPC effects of RO might be related to its regulatory impact via the PI3K-AKT signaling pathway, the Wnt signaling pathway, and the cAMP signaling pathway by targeting VEGFA, TP53, and HSPA8. The findings of potential key targets may provide new clues for NPC's treatments with the RO adjunctive therapy.


The Effect of 400 µg Inhaled Salbutamol on 3 km Time Trial Performance in a Low Humidity Environment.
The Objectives of the study were to investigate whether 400 µg inhaled salbutamol influences 3 km running time-trial performance and lung function in eucapnic voluntary hyperpnoea positive (EVH+ve) and negative (EVH-ve) individuals. Fourteen male participants (22.4 ± 1.6yrs; 76.4 ± 8.7kg; 1.80 ± 0.07 m); (7 EVH+ve; 7 EVH-ve) were recruited following written informed consent. All participants undertook an EVH challenge to identify either EVH+ve (↓FEV1>10%) or EVH-ve (↓FEV1<10%). Participants performed three separate 3 km running time-trials in a low-humidity (20-25%) environment on a non-motorized treadmill, 15 minutes following inhalation of salbutamol (400 µg), placebo (non-active inhalant) or control (no inhalant), in a randomized, single-blind, repeated measures design. Forced vital capacity maneuvers were performed at baseline, 10 minutes post inhalation and post time-trial. Time to complete 3 km and lung function data were analyzed using mixed model repeated measures ANOVA. Significance was assumed at p < 0.05. All EVH+ve participants had FEV1 falls from baseline between 10-25% post-challenge. There was no difference in performance time between trial conditions in EVH+ve (1012.7 ± 129.6s; 1002.4 ± 123.1s; 1015.9 ± 113.0s) (p = 0.774) and EVH-ve (962.1 ± 99.2s; 962.0 ± 76.2s; 950.8 ± 84.9s) (p = 0.401) groups for salbutamol, placebo and control trials, respectively. Exercising heart rate was significantly higher (p = 0.05) in the salbutamol trial (183 ± 8 beatsˑmin-1) compared to control (180 ± 9 beatsˑmin-1) with a trend towards significance (p=0.06) in the placebo trial (179 ± 9 beatsˑmin-1) for the pooled groups, no differences were seen between trials in groups individually. There was an increase in FEV1 in both EVH+ve (4.01 ± 0.8L; 4.26 ± 0.7L; 4.25 ± 0.5L) and EVH-ve (4.81 ± 0.4L; 5.1 ± 0.4L; 5.1 ± 0.5L) groups which was significant post-inhalation (p = 0.01; p = 0.02), but not post-time-trial (p = 0.27; p = 0.06), respectively, following salbutamol. EVH+ve participants did not demonstrate significant falls (>10% from baseline) in FEV1 following any time-trial. Administration of 400µg inhaled salbutamol does not improve 3 km time-trial performance in either mild EVH+ve or EVH-ve individuals despite significantly increased HR and FEV1.


Low 2-dimensional CD4 T cell receptor affinity for myelin sets in motion delayed response kinetics.
T cells recognizing self-peptides that mediate autoimmune disease and those that are responsible for efficacious immunity against pathogens may differ in affinity for antigen due to central and peripheral tolerance mechanisms. Here we utilize prototypical self-reactive (myelin) and viral-specific (LCMV) T cells from T cell receptor (TCR) transgenic mice (2D2 and SMARTA, respectively) to explore affinity differences. The T cells responsive to virus possessed >10,000 fold higher 2D affinity as compared to the self-reactive T cells. Despite their dramatically lower affinity for their cognate ligand, 2D2 T cells respond with complete, albeit delayed, activation (proliferation and cytokine production). SMARTA activation occurs rapidly, achieving peak phosphorylation of p38 (1 minute), Erk (30 minutes), and Jun (3 hours) as well as CD69 and CD25 upregulation (3 and 6 hours, respectively), with a corresponding early initiation of proliferation. 2D2 stimulation with MOG results in altered signaling--no phospho-Erk or phospho-p38 accumulation, significantly delayed activation kinetics of Jun (12 hours), and delayed but sustained SHP-1 activity--as well as delayed CD69 and CD25 expression (12-24 hours), and slow initiation of proliferation. This delay was not intrinsic to the 2D2 T cells, as a more potent antigen with >100-fold increased 2D affinity restored rapid response kinetics in line with those identified for the viral antigen. Taken together, these data demonstrate that time can offset low TCR affinity to attain full activation and suggest a mechanism by which low affinity T cells participate in autoimmune disease.


Annulate in four cases of diffuse lymphocytic lymphoma.
Annulate lamellae were observed in 4 cases of diffuse lymphocytic lymphoma. Two lymphomas were well differentiated and 2 were of intermediate differentiation. These lymphomas had additional abnormalities of membranes: specifically, excessive blebbing of nuclear membranes and lattice-like formations of tubular endoplasmic reticulum.


Verrucous carcinoma of the vulva in a patient infected with the human immunodeficiency virus.
Verrucous carcinomas of the vulva are rare and have not been reported in women infected with the human immunodeficiency virus. We present such a case in a 32-year-old woman characterized by bladder involvement that failed therapy with 13-cis-retinoic acid and interferon-alpha and required anterior exenteration.


Impact of Gut and Metabolic Hormones on Feeding Reward.
Ingestion of food activates a cascade of endocrine responses (thereby reflecting a contemporaneous feeding status) that include the release of hormones from the gastrointestinal (GI) tract, such as cholecystokinin (CCK), glucagonlike peptide YY (PYY), peptide PP, and oleoylethanolamide, as well as suppression of ghrelin secretion. The pancreas and adipose tissue, on the other hand, release hormones that serve as a measure of the current metabolic state or the long-term energy stores, that is, insulin, leptin, and adiponectin. It is well known and intuitively understandable that these hormones target either directly (by crossing the blood-brain barrier) or indirectly (e.g., via vagal input) the "homeostatic" brainstem-hypothalamic pathways involved in the regulation of appetite. The current article focuses on yet another target of the metabolic and GI hormones that is critical in inducing changes in food intake, namely, the reward system. We discuss the physiological basis of this functional interaction, its importance in the control of appetite, and the impact that disruption of this crosstalk has on energy intake in select physiological and pathophysiological states. We conclude that metabolic and GI hormones have a capacity to strengthen or weaken a response of the reward system to a given food, and thus, they are fundamental in ensuring that feeding reward is plastic and dependent on the energy status of the organism. © 2021 American Physiological Society. Compr Physiol 11:1425-1447, 2021.


Tobacco smoking and risk of endometriosis: a systematic review and meta-analysis.
Since conflicting results have been published on the role of tobacco smoking on the risk of endometriosis, we provide an up-to-date summary quantification of this potential association. We performed a PubMed/MEDLINE search of the relevant publications up to September 2014, considering studies on humans published in English. We searched the reference list of the identified papers to find other relevant publications. Case-control as well as cohort studies have been included reporting risk estimates on the association between tobacco smoking and endometriosis. 38 of the 1758 screened papers met the inclusion criteria. The selected studies included a total of 13,129 women diagnosed with endometriosis. Academic hospitals. Risk of endometriosis in tobacco smokers. We obtained the summary estimates of the relative risk (RR) using the random effect model, and assessed the heterogeneity among studies using the χ(2) test and quantified it using the I(2) statistic. As compared to never-smokers, the summary RR were 0.96 (95% CI 0.86 to 1.08) for ever smokers, 0.95 (95% CI 0.81 to 1.11) for former smokers, 0.92 (95% CI 0.82 to 1.04) for current smokers, 0.87 (95% CI 0.70 to 1.07) for moderate smokers and 0.93 (95% CI 0.69 to 1.26) for heavy smokers. The present meta-analysis provided no evidence for an association between tobacco smoking and the risk of endometriosis. The results were consistent considering ever, former, current, moderate and heavy smokers, and across type of endometriosis and study design.


Steroid-receptor fusion of the human immunodeficiency virus type 1 Rev transactivator: mapping cryptic functions of the arginine-rich motif.
The human immunodeficiency virus type 1 (HIV-1) transactivator Rev is a nuclear protein that regulates expression of certain HIV-1 transcripts by binding to an RNA target element (the RRE) present in these transcripts. A short arginine-rich sequence in Rev contains the signals required to direct this protein into nuclei, where it associates preferentially with nucleoli. We created a steroid-inducible transactivator by fusing Rev with the steroid-binding domain of the glucocorticoid receptor (GR). This Rev/GR protein remains inactive in the cytoplasm when steroids are absent, but it enters the nucleus and initiates transactivation within minutes after exposure to dexamethasone. Although the GR moiety is sufficient to transport Rev/GR into nuclei, mutation of certain residues in the arginine-rich region blocks nucleolar localization and also inhibits transactivation. We find that other mutations in this region, however, can abolish the function of Rev/GR without affecting its localization; the latter phenotype may reflect a specific defect in binding of the RRE.


Functional Effects In Silico Prediction for Androgen Receptor Ligand-Binding Domain Novel I836S Mutation.
Over 1000 mutations are described in the androgen receptor (AR) gene. Of those, about 600 were found in androgen insensitivity syndrome (AIS) patients, among which 400 mutations affect the ligand-binding domain (LBD) of the AR protein. Recently, we reported a novel missense mutation c.2507T>G I836S (ClinVarID: 974911) in a patient with complete AIS (CAIS) phenotype. In the present study, we applied a set of computational approaches for the structural analysis of the ligand-binding domains in a wild-type and mutant AR to evaluate the functional impact of the novel I836S mutation. We revealed that the novel I836S substitution leads to a shorter existence time of the ligand's gating tunnel and internal cavity, occurring only in the presence of S836 phosphorylation. Additionally, the analysis of phosphorylation of the 836 mutant residues explained the negative impact on AR homodimerization, since monomer surface changes indirectly impacted the binding site. Our analyses provide evidence that I836S causes disruptions of AR protein functionality and development of CAIS clinical features in patients.


Effect of Spectral Contrast Enhancement on Speech-on-Speech Intelligibility and Voice Cue Sensitivity in Cochlear Implant Users.
Speech intelligibility in the presence of a competing talker (speech-on-speech; SoS) presents more difficulties for cochlear implant (CI) users compared with normal-hearing listeners. A recent study implied that these difficulties may be related to CI users' low sensitivity to two fundamental voice cues, namely, the fundamental frequency (F0) and the vocal tract length (VTL) of the speaker. Because of the limited spectral resolution in the implant, important spectral cues carrying F0 and VTL information are expected to be distorted. This study aims to address two questions: (1) whether spectral contrast enhancement (SCE), previously shown to enhance CI users' speech intelligibility in the presence of steady state background noise, could also improve CI users' SoS intelligibility, and (2) whether such improvements in SoS from SCE processing are due to enhancements in CI users' sensitivity to F0 and VTL differences between the competing talkers. The effect of SCE on SoS intelligibility and comprehension was measured in two separate tasks in a sample of 14 CI users with Cochlear devices. In the first task, the CI users were asked to repeat the sentence spoken by the target speaker in the presence of a single competing talker. The competing talker was the same target speaker whose F0 and VTL were parametrically manipulated to obtain the different experimental conditions. SoS intelligibility, in terms of the percentage of correctly repeated words from the target sentence, was assessed using the standard advanced combination encoder (ACE) strategy and SCE for each voice condition. In the second task, SoS comprehension accuracy and response times were measured using the same experimental setup as in the first task, but with a different corpus. In the final task, CI users' sensitivity to F0 and VTL differences were measured for the ACE and SCE strategies. The benefit in F0 and VTL discrimination from SCE processing was evaluated with respect to the improvement in SoS perception from SCE. While SCE demonstrated the potential of improving SoS intelligibility in CI users, this effect appeared to stem from SCE improving the overall signal to noise ratio in SoS rather than improving the sensitivity to the underlying F0 and VTL differences. A second key finding of this study was that, contrary to what has been observed in a previous study for childlike voice manipulations, F0 and VTL manipulations of a reference female speaker (target speaker) toward male-like voices provided a small but significant release from masking for the CI users tested. The present findings, together with those previously reported in the literature, indicate that SCE could serve as a possible background-noise-reduction strategy in commercial CI speech processors that could enhance speech intelligibility especially in the presence of background talkers that have longer VTLs compared with the target speaker.


Dominant negative alpha-subunit of FTase inhibits effects of insulin and IGF-I in MCF-7 cells.
We recently designed a dominant negative (DN) farnesyltransferase (FTase)/geranyl-gerahyltransferase I (GGTase I) alpha-subunit that when expressed in vascular smooth muscle cells decreased insulin-stimulated phosphorylation of FTase, FTase activity, amounts of farnesylated p21Ras, DNA synthesis, and cell migration. Currently, we explored the inhibitory effects of DN FTase/GGTase I alpha-subunit in MCF-7 cells on IGF-1- and insulin-stimulated DNA synthesis and cell proliferation. Expression of the DN FTase/GGTase I alpha-subunit completely blocked IGF-1- and insulin-stimulated BrdU incorporation and cell count. DN FTase/GGTase I alpha-subunit inhibited insulin-stimulated phosphorylation of FTase/GGTase I alpha-subunit, FTase and GGTase I activity, and prenylation of p21Ras and RhoA. Expression of DN FTase/GGTase I alpha-subunit diminished IGF-1- and insulin-stimulated phosphorylation of ERK (extracellular signal-regulated kinase), but had no effect on IGF-1- and insulin-stimulated phosphorylation of Akt. Taken together, these data suggest that DN FTase/GGTase I alpha-subunit can assuage the mitogenic effects of IGF-1 and insulin on MCF-7 breast cancer cells.


Solvent-Assisted Fractionation of Oligomeric Cellulose and Reversible Transformation of Cellulose II and IV.
Oligomeric cellulose with an average degree of polymerization of 7.68 and a polydispersity of 1.04 has been fractionated using solution processes. Three fractions have been obtained through initial dissolution, subsequent crystallization, and solvent precipitation, respectively. The resulting oligocellulose fraction has an average degree of polymerization of 7.70 and a polydispersity of 1.01, respectively. Cellulose IV2 crystals form in the oligocellulose fraction, and reversibly transform to II and back to IV using simple solvents.


Impact of the supramolecular structure of cellulose on the efficiency of enzymatic hydrolysis.
The efficiency of enzymatic hydrolysis is reduced by the structural properties of cellulose. Although efforts have been made to explain the mechanism of enzymatic hydrolysis of cellulose by considering the interaction of cellulolytic enzymes with cellulose or the changes in the structure of cellulose during enzymatic hydrolysis, the process of cellulose hydrolysis is not yet fully understood. We have analysed the characteristics of the complex supramolecular structure of cellulose on the nanometre scale in terms of the spatial distribution of fibrils and fibril aggregates, the accessible surface area and the crystallinity during enzymatic hydrolysis. Influence of the porosity of the substrates and the hydrolysability was also investigated. All cellulosic substrates used in this study contained more than 96% cellulose. Conversion yields of six cellulosic substrates were as follows, in descending order: nano-crystalline cellulose produced from never-dried soda pulp (NCC-OPHS-ND) > never-dried soda pulp (OPHS-ND) > dried soda pulp (OPHS-D) > Avicel > cotton treated with sodium hydroxide (cotton + NaOH) > cotton. No significant correlations were observed between the yield of conversion and supramolecular characteristics, such as specific surface area (SSA) and lateral fibril dimensions (LFD). A strong correlation was found between the average pore size of the starting material and the enzymatic conversion yield. The degree of crystallinity was maintained during enzymatic hydrolysis of the cellulosic substrates, contradicting previous explanations of the increasing crystallinity of cellulose during enzymatic hydrolysis. Both acid and enzymatic hydrolysis can increase the LFD, but no plausible mechanisms could be identified. The sample with the highest initial degree of crystallinity, NCC-OPHS-ND, exhibited the highest conversion yield, but this was not accompanied by any change in LFD, indicating that the hydrolysis mechanism is not based on lateral erosion.


FDG PET Data is Associated with Cognitive Performance in Patients from a Memory Clinic.
Various reasons may lead to cognitive symptoms in elderly, including the development of cognitive decline and dementia. Often, mixed pathologies such as neurodegeneration and cerebrovascular disease co-exist in these patients. Diagnostic work-up commonly includes imaging modalities such as FDG PET, MRI, and CT, each delivering specific information. To study the informative value of neuroimaging-based data supposed to reflect neurodegeneration (FDG PET), cerebral small vessel disease (MRI), and cerebral large vessel atherosclerosis (CT) with regard to cognitive performance in patients presenting to our memory clinic. Non-parametric partial correlations and an ordinal logistic regression model were run to determine relationships between scores for cortical hypometabolism, white matter hyperintensities, calcified plaque burden, and results from Mini-Mental State Examination (MMSE). The final study group consisted of 162 patients (female: 94; MMSE: 6-30). Only FDG PET data was linked to and predicted cognitive performance (r(157) = -0.388, p < 0.001). Overall, parameters linked to cerebral small and large vessel disease showed no significant association with cognition. Further findings demonstrated a relationship between white matter hyperintensities and FDG PET data (r(157) = 0.230, p = 0.004). Only FDG PET imaging mirrors cognitive performance, presumably due to the examination's ability to reflect neurodegeneration and vascular dysfunction, thus capturing a broader spectrum of pathologies. This makes the examination a useful imaging-based diagnostic tool in the work-up of patients presenting to a memory clinic. Parameters of vascular dysfunction alone as depicted by conventional MRI and CT are less adequate in such a situation, most likely because they reflect one pathology complex only.