Ligand recognition by influenza virus. The binding of bivalent sialosides.
Infection by influenza virus is initiated by a cellular adhesion event that is mediated by the viral protein, hemagglutinin, which is exposed on the surface of the virion. Hemagglutinin recognizes and binds to cell surface sialic acid residues. Although each individual ligand binding interaction is weak, the high affinity of influenza virus for cells that bear sialic acid residues is thought to result from a multivalent attachment process involving many similar recognition events. To evaluate such binding we have synthesized three series of compounds, each containing two sialic acid residues separated by spacers of different length, and have tested them as ligands for influenza hemagglutinin. No increased binding to the bromelain-released hemagglutinin ectodomain was seen for any of the bivalent compounds as determined by 1H NMR titration. In contrast, however, a spacer length between sialic acid residues of approximately 55 A sharply increases the binding of these bidentate species to whole virus as determined by hemagglutination inhibition assays. The most effective compound containing glycines in the linking chain displayed 100-fold increased affinity for whole virus over the paradigm monovalent ligand, Neu5Ac alpha 2Me.


Prediction of GFR in liver transplant candidates.
Kidney function frequently is impaired in patients with cirrhosis; however, glomerular filtration rate (GFR) is difficult to estimate in these patients by using standard clinical markers. The aim of our study is to compare GFR calculated from renal clearance of iodine 125-labeled iothalamate ((125)I-iothalamate) with the plasma decay technique and the Modification of Diet in Renal Disease (MDRD) and Cockroft-Gault (CG) prediction equations. We performed a cross-sectional study of patients with liver cirrhosis being evaluated for transplantation (50% Child's class C); 89% had ascites or edema and 44% were men aged 55 +/- 2 years. Average pretest blood urea nitrogen level was 16 +/- 2 mg/dL (5.7 +/- 0.7 mmol/L); serum creatinine, 1.0 +/- 0.1 mg/dL (88 +/- 9 micromol/L; range, 0.6 to 1.7 mg/dL [53 to 150 micromol/L]); plasma albumin, 3.14 +/- 0.16 g/dL (31.4 +/- 1.6 g/L); and total bilirubin, 4.0 +/- 0.7 mg/dL (67 +/- 11.3 micromol/L). Kidney function was measured by means of simultaneous plasma and renal clearance of (125)I-iothalamate (Glofil-125; Cypros Pharmaceutical Corp, Carlsbad, CA) and the MDRD and CG equations. GFRs were 58.2 +/- 5.1 mL/min/1.73 m(2) by renal clearance of (125)I-iothalamate and 76.7 +/- 7.2 mL/min/1.73 m(2) by the plasma decay technique (+18.5 mL/min, or 32%; P = 0.0004). GFR by the MDRD equation was 76.9 +/- 7.8 mL/min/1.73 m(2) (+18.7 mL/min, or 32%; P = 0.0004 versus renal iothalamate; r(2) = 0.57). GFR by the CG equation was the least accurate (+30.1 mL/min, or 52%; P = 0.0001 versus renal iothalamate). The current clinically used CG and MDRD equations to estimate kidney function in patients with cirrhosis and volume excess and the (125)I-iothalamate plasma decay technique are inaccurate because they overestimate GFR. It seems very unlikely that accurate and reliable formulas will be developed that are able to replace the formal measurement of GFR in patients with liver cirrhosis. Therefore, we conclude that despite the additional complexity, renal clearance techniques should be used to assess GFR accurately in patients with liver cirrhosis and ascites.


Is interspecies hydrogen transfer needed for toluene degradation under sulfate-reducing conditions?
Sediments from a hydrocarbon-contaminated aquifer, where periodic shifts between sulfate reduction and methanogenesis occurred, were examined to determine whether the degradation of toluene under sulfate-reducing conditions depended on interspecies hydrogen transfer. Toluene degradation under sulfate-reducing conditions was inhibited by the addition of 5 mM sodium molybdate, but the activity was not restored upon the addition of an actively growing, hydrogen-using methanogen. Toluene degradation was not inhibited in microcosms where hydrogen levels were maintained at a level theoretically sufficient to inhibit toluene degradation if the process proceeded via interspecies hydrogen transfer. Finally, the addition of carbon monoxide, a potent inhibitor of hydrogenase activity, inhibited hydrogen but not toluene consumption in sulfate-reducing microcosms. These results suggest that toluene is degraded directly by sulfate-reducing bacteria without the involvement of interspecies hydrogen transfer. The sequence of experiments used to reach this conclusion could be applied to determine the role of interspecies hydrogen transfer in the degradation of a variety of compounds in different environments or under different terminal electron-accepting conditions.


A comparison of vowel production by children with multichannel cochlear implants or tactile aids: perceptual evidence.
To examine changes in perceived vowel production accuracy over time in prelingually deafened children who use a multichannel cochlear implant (Nucleus 22 channel) or a multichannel tactile aid (Tactaid 7) and to compare the levels of perceived vowel production accuracy attained by the two device groups. The subjects were participants in longitudinal studies of the effects of sensory aids on the development of perceptual, speech, and language skills. As part of these studies, imitative vowel productions were elicited and transcribed before each child received their sensory aid and at 6 mo intervals thereafter. Data for the current study was obtained from the predevice interval and a later interval (postdevice) that was an average of 1.8 yr after the subjects received their respective devices. The subjects' vowel productions were judged for accuracy in matching an imitative model and for correctness of vowel features (height and place). Within-group analyses were completed to determine if vowel production scores improved over the course of the study for each device group. Between group comparisons were performed to examine differences in mean scores at each interval. Before receiving their multichannel devices, the two groups of subjects demonstrated similar imitative vowel production skills. After an average of 1.8 yr of device use, the cochlear implant subjects demonstrated significantly improved production of diphthongs and all vowel categories except low vowels. The Tactaid 7 subjects demonstrated significant improvement only in the production of diphthongs. Thus, cochlear implant recipients' vowel production skills were found to be significantly better than those of the Tactaid 7 users after a comparable amount of device experience. The vowel production gains of the cochlear implant subjects were similar in amount to those noted in other studies of children who use the Nucleus multichannel cochlear implant and further confirm the potential of this device for improved speech production in prelingually deafened children. The differences between the performance of the two groups demonstrate that vowel production skills improved to a greater degree through use of a multichannel cochlear implant than through use of the Tactaid 7.


Quality of Life, Perception of Disease and Coping Strategies in Patients with Hemophilia in Spain and El Salvador: A Comparative Study.
Hemophilia is characterized by the development of joint bleeds that cause long-term joint damage (hemophilic arthropathy). Joint damage leads to disability and affects psychosocial aspects in patients with hemophilia. To compare the clinical situation, perception of disease and quality of life, and coping strategies in adult patients with hemophilia in El Salvador and Spain. In this comparative clinical study, 43 patients with hemophilia aged between 18 and 50 years old from Spain and El Salvador participated. After obtaining the patients' consent, they completed the Illness Perception Questionnaire-Revised (IPQ-R), Hemophilia-QoL and Inventory of Coping strategies questionnaires. Joint status was assessed using the Hemophilia Joint Health Score and based on a record of clinical and treatment data. Hemophilia patients from Spain showed an improved perception of quality of life (p <0.05), although there were only differences in the self-criticism variable (p = 0.04) for coping strategies. Joint damage and age correlated (p <0.05) negatively with perception of disease, perceived quality of life and coping strategies in both populations. There were differences (p <0.05) between the two populations based on HIV and HCV coinfections in perception of disease and perceived quality of life. Patients with hemophilia in El Salvador exhibit a poorer perception of disease and quality of life. Despite differences in access to treatment from one country to the other, there is no difference in coping with the disease. Older patients are better able to adapt to the disease.


Detection of esophageal varices using CT and MRI.
The development of esophageal varices in cirrhotic patients carries a significant risk of hemorrhage and associated morbidity/mortality. Universal endoscopic screening, however, is invasive and expensive. Conversely, cirrhotic patients often have imaging findings which suggest portal hypertension. The aim of this study was to evaluate the ability of CT and/or MRI to detect esophageal varices compared to EGD. Medical records from 2000 to 2007 were retrospectively reviewed. CT and/or MRI images were included if performed within 90 days of EGD. Two blinded, experienced radiologists were asked to review images for the presence of esophageal varices, as well as other findings associated with portal hypertension. Sensitivity, specificity, PPV and NPV were calculated using EGD findings as the gold standard. A total of 195 patients and 142 patients met criteria for CT and MRI, respectively. The sensitivity of CT to detect EGD varices was 58-89%, but increased to 65-100% when specifically looking at large endoscopic varices. Overall specificity was 68-82%, but increased to 97-100% when applying ≥4 mm varices criteria. CT was superior to MRI in the detection of endoscopic varices; the addition of other portal hypertension stigmata did not improve results. The exclusion of large endoscopic varices by CT, using standardized criteria, may obviate the need or frequency of EGD screening in select patient populations. Alternatively, CT findings highly suggestive of esophageal varices in cirrhotic patients may warrant further investigation and/or treatment. Further studies are needed to validate these findings.


Usefulness and Potential Pitfalls of Long-Acting Growth Hormone Analogs.
Daily recombinant human GH (rhGH) is currently approved for use in children and adults with GH deficiency (GHD) in many countries with relatively few side-effects. Nevertheless, daily injections can be painful and distressing for some patients, often resulting in non-adherence and reduction of treatment outcomes. This has prompted the development of numerous long-acting GH (LAGH) analogs that allow for decreased injection frequency, ranging from weekly, bi-weekly to monthly. These LAGH analogs are attractive as they may theoretically offer increased patient acceptance, tolerability, and therapeutic flexibility. Conversely, there may also be pitfalls to these LAGH analogs, including an unphysiological GH profile and differing molecular structures that pose potential clinical issues in terms of dose initiation, therapeutic monitoring, incidence and duration of side-effects, and long-term safety. Furthermore, fluctuations of peak and trough serum GH and IGF-I levels and variations in therapeutic efficacy may depend on the technology used to prolong GH action. Previous studies of some LAGH analogs have demonstrated non-inferiority compared to daily rhGH in terms of increased growth velocity and improved body composition in children and adults with GHD, respectively, with no significant unanticipated adverse events. Currently, two LAGH analogs are marketed in Asia, one recently approved in the United States, another previously approved but not marketed in Europe, and several others proceeding through various stages of clinical development. Nevertheless, several practical questions still remain, including possible differences in dose initiation between naïve and switch-over patients, methodology of dose adjustment/s, timing of measuring serum IGF-I levels, safety, durability of efficacy and cost-effectiveness. Long-term surveillance of safety and efficacy of LAGH analogs are needed to answer these important questions.


Community factors affecting participation in larval source management for malaria control in Chikwawa District, Southern Malawi.
To further reduce malaria, larval source management (LSM) is proposed as a complementary strategy to the existing strategies. LSM has potential to control insecticide resistant, outdoor biting and outdoor resting vectors. Concerns about costs and operational feasibility of implementation of LSM at large scale are among the reasons the strategy is not utilized in many African countries. Involving communities in LSM could increase intervention coverage, reduce costs of implementation and improve sustainability of operations. Community acceptance and participation in community-led LSM depends on a number of factors. These factors were explored under the Majete Malaria Project in Chikwawa district, southern Malawi. Separate focus group discussions (FGDs) were conducted with members from the general community (n = 3); health animators (HAs) (n = 3); and LSM committee members (n = 3). In-depth interviews (IDIs) were conducted with community members. Framework analysis was employed to determine the factors contributing to community acceptance and participation in the locally-driven intervention. Nine FGDs and 24 IDIs were held, involving 87 members of the community. Widespread knowledge of malaria as a health problem, its mode of transmission, mosquito larval habitats and mosquito control was recorded. High awareness of an association between creation of larval habitats and malaria transmission was reported. Perception of LSM as a tool for malaria control was high. The use of a microbial larvicide as a form of LSM was perceived as both safe and effective. However, actual participation in LSM by the different interviewee groups varied. Labour-intensiveness and time requirements of the LSM activities, lack of financial incentives, and concern about health risks when wading in water bodies contributed to lower participation. Community involvement in LSM increased local awareness of malaria as a health problem, its risk factors and control strategies. However, community participation varied among the respondent groups, with labour and time demands of the activities, and lack of incentives, contributing to reduced participation. Innovative tools that can reduce the labour and time demands could improve community participation in the activities. Further studies are required to investigate the forms and modes of delivery of incentives in operational community-driven LSM interventions.


Biology, prognosis and response to therapy of breast carcinomas according to HER2 score.
The standardization of the HER2 score and recent changes in therapeutic modalities points to the need for a reevaluation of the role of HER2 in recently diagnosed breast carcinoma. A multicenter, retrospective study of 1794 primary breast carcinomas diagnosed in Italy in 2000/2001 and scored in HER2 four categories according to immunohistochemistry was conducted. Ductal histotype, vascular invasion, grade, MIB1 positivity, estrogen and progesterone receptor expression differed significantly in HER2 3+ tumors compared with the other categories. HER2 2+ tumors almost showed values intermediate between those of the negative and the 3+ subgroups. The characteristics of HER2 1+ tumors were found to be in between those of HER2 0 and 2+ tumors. With a median follow-up of 54 months, HER2 3+ status was associated with higher relapse rates in node-positive and node-negative subgroups, while HER2 2+ only in node positive. Analysis of relapses according to type of therapy provided evidence of responsiveness of HER2-positive tumors to chemotherapy, especially taxanes. The present prognostic significance of HER2 is correlated to receptor expression level and points to the need to consider HER2 2+ and HER2 3+ tumors as distinct diseases with different outcomes and specific features.


Blockage of the upregulation of voltage-gated sodium channel nav1.3 improves outcomes after experimental traumatic brain injury.
Excessive active voltage-gated sodium channels are responsible for the cellular abnormalities associated with secondary brain injury following traumatic brain injury (TBI). We previously presented evidence that significant upregulation of Nav1.3 expression occurs in the rat cortex at 2 h and 12 h post-TBI and is correlated with TBI severity. In our current study, we tested the hypothesis that blocking upregulation of Nav1.3 expression in vivo in the acute stage post-TBI attenuates the secondary brain injury associated with TBI. We administered either antisense oligodeoxynucleotides (ODN) targeting Nav1.3 or artificial cerebrospinal fluid (aCSF) at 2 h, 4 h, 6 h, and 8 h following TBI. Control sham animals received aCSF administration at the same time points. At 12 h post-TBI, Nav1.3 messenger ribonucleic acid (mRNA) levels in bilateral hippocampi of the aCSF group were significantly elevated, compared with the sham and ODN groups (p<0.01). However, the Nav1.3 mRNA levels in the uninjured contralateral hippocampus of the ODN group were significantly lowered, compared with the sham group (p<0.01). Treatment with antisense ODN significantly decreased the number of degenerating neurons in the ipsilateral hippocampal CA3 and hilar region (p<0.01). A set of left-to-right ratio value analyzed by magnetic resonance imaging T2 image on one day, three days, and seven days post-TBI showed marked edema in the ipsilateral hemisphere of the aCSF group, compared with that of the ODN group (p<0.05). The Morris water maze memory retention test showed that both the aCSF and ODN groups took longer to find a hidden platform, compared with the sham group (p<0.01). However, latency in the aCSF group was significantly higher than in the ODN group (p<0.05). Our in vivo Nav1.3 inhibition studies suggest that therapeutic strategies to block upregulation of Nav1.3 expression in the brain may improve outcomes following TBI.


Adaptation and validation of the Italian Pediatric Voice Handicap Index.
To evaluate the internal consistency, reliability and clinical validity of the Italian version of the Pediatric Voice Handicap Index (pVHI). The parents of 30 children with dysphonia and 43 asymptomatic children were included in the study. Each parent was asked to complete the Italian pVHI autonomously. The voice of each child was assessed perceptually through the GRB parameters of the GRBAS scale. Internal consistency was analyzed through Cronbach's α coefficient. For test-retest reliability analysis, the Italian pVHI was filled twice, with a 2-week interval, and the scores obtained were compared through the Pearson correlation test. Clinical validity was assessed comparing the scores obtained in the pathological and the control group using the Mann-Whitney test. Finally, the correlation between pVHI and the perceptual parameters was assessed. All of the parents filled in the entire questionnaire autonomously. An optimal internal consistency was found (α = 0.95); the test-retest reliability in the parents of both groups of children was high (r > 0.88). The control group scored significantly lower than the pathological group (p = 0.0001). The pVHI scores positively correlated with perceptual assessment of voice disorders. The Italian pVHI is easily administered, highly reproducible, and exhibits excellent clinical validity.


Effect of oestradiol-17beta and progesterone on the metabolism of [1,2(-3)H] progesterone by the rabbit endometrium and myometrium.
The metabolism of [3H]progesterone in the rabbit endometrium and myometrium was studied in vitro. The major metabolities identified were 5alpha-pregnane-3,20-dione, 20alpha-hydroxypregn-4-en-3-one, 3beta-hydroxy-5alpha-preganan-20-one and 5alpha-pregnane-3beta,20alpha-diol. Other minor metabolites tentatively identified were 3alpha-hydroxy-5beta-pregnan-20-one,20alpha-hydroxy-5beta-pregnan-3-one and 5beta-pregnane-3alpha,20alpha-diol. The ability of the endometrium to metabolize progesterone on a unit weight bais was about 2.7 times that of the myometrium. The metabolism of [3H]progesterone in the rabbit uterus under the influnce of oestradiol-17beta and progesterone was studied. The ability of the oestradiol-treated rabbit uterus to metabolize progesterone was increased to 3.47 times that of the overiectomized control uterus, whereas the oestradiol-progesterone-treated rabbit uterus metabolized only 1.86 times that of the control. Study of the metabolism of progesterone with uterine subcellular preparations revealed that the 5alpha-reductase enzyme was present mainly in the nuclear fraction; 20alpha-hydroxysteroid dehydrogenase was found in the cytosol fraction and 3beta-hydroxysteroid dehydrogenase in the particulate fraction of the uterus. The metabolic pathways of progesterone in the rabbit uterine tissue are discussed.


Deoxyribonucleic acid synthesis in bacteriophage SPO1-infected Bacillus subtilis. I. Bacteriophage deoxyribonucleic acid synthesis and fate of host deoxyribonucleic acid in normal and polymerase-deficient strains.
The effect of bacteriophage SPO1 infection of Bacillus subtilis and a deoxyribonucleic acid (DNA) polymerase-deficient (pol(-)) mutant of this microorganism on the synthesis of DNA has been examined. Soon after infection, the incorporation of deoxyribonucleoside triphosphates into acid-insoluble material by cell lysates was greatly reduced. This inhibition of host DNA synthesis was not a result of host chromosome degradation nor did it appear to be due to the induction of thymidine triphosphate nucleotidohydrolase. Examination of the host chromosome for genetic linkage throughout the lytic cycle indicated that no extensive degradation occurred. After the inhibition of host DNA synthesis, a new polymerase activity arose which directed the synthesis of phage DNA. This new activity required deoxyribonucleoside triphosphates as substrates, Mg(2+) ions, and a sulfhydryl reducing agent, and it was stimulated in the presence of adenosine triphosphate. The phage DNA polymerase, like that of its host, was associated with a fast-sedimenting cell membrane complex. The pol(-) mutation had no effect on the synthesis of phage DNA or production of mature phage particles.


ATP Mediates Phase Separation of Disordered Basic Proteins by Bridging Intermolecular Interaction Networks.
ATP is an abundant molecule with crucial cellular roles as the energy currency and a building block of nucleic acids and for protein phosphorylation. Here we show that ATP mediates the phase separation of basic intrinsically disordered proteins (bIDPs). In the resulting condensates, ATP is highly concentrated (apparent partition coefficients at 200-5000) and serves as bridges between bIDP chains. These liquid-like droplets have some of the lowest interfacial tension (~25 pN/μm) but high zero-shear viscosities (1-15 Pa s) due to the bridged protein networks, and yet their fusion has some of the highest speeds (~1 μm/ms). The rapid fusion manifests extreme shear thinning, where the apparent viscosity is lower than zero-shear viscosity by over 100-fold, made possible by fast reformation of the ATP bridges. At still higher concentrations, ATP does not dissolve bIDP droplets but results in aggregates and fibrils.


Characterization of a C5a receptor on human polymorphonuclear leukocytes (PMN).
Elucidation of the interactions between C5a and granulocytes is central to understanding the role of C5a in inflammation. In this study, interactions between C5a and PMN have been studied at two levels. Binding of human C5a to intact human cells has been characterized by using the radiolabeled ligand 125I-C5a. Binding is shown to be reversible, saturable, and to reach equilibrium in 60 to 90 min at 0 degrees C. Results show high affinity C5a binding sites with Kd = 2 X 10(-9) M and a range of 50,000 to 113,000 binding sites per PMN. These values for C5a receptors are comparable with the number of fMLP and LTB4 receptors on PMN. Binding of C5a to PMN fails to reach equilibrium at 37 degrees C because there is an irreversible loss of available surface receptors caused by an active internalization of the ligand-receptor complex. Interactions between C5a and human PMN were characterized further by cross-linking experiments, with the use of ethylene glycol bis succinimidylsuccinate (EGS). Cross-linking of 125I-C5a to intact PMN followed by subcellular fractionation revealed a single radioactive band present only in the plasma membrane fraction and visualized by autoradiography. Similar experiments resulted in a covalent linkage between 125I-C5a and a component in the isolated plasma membrane of PMN. The covalent complex containing C5a and a putative receptor has been visualized by autoradiography as a single 60,000 Mr complex on SDS-PAGE. The complex is not present when experiments are performed in the presence of excess unlabeled C5a or in the absence of EGS. Therefore, the putative receptor for C5a on human neutrophils is estimated to be approximately 48,000 Mr, assuming contribution of 12,000 to 13,000 daltons by the ligand 125I-C5a.


Enzalutamide with Standard First-Line Therapy in Metastatic Prostate Cancer.
Enzalutamide, an androgen-receptor inhibitor, has been associated with improved overall survival in men with castration-resistant prostate cancer. It is not known whether adding enzalutamide to testosterone suppression, with or without early docetaxel, will improve survival in men with metastatic, hormone-sensitive prostate cancer. In this open-label, randomized, phase 3 trial, we assigned patients to receive testosterone suppression plus either open-label enzalutamide or a standard nonsteroidal antiandrogen therapy (standard-care group). The primary end point was overall survival. Secondary end points included progression-free survival as determined by the prostate-specific antigen (PSA) level, clinical progression-free survival, and adverse events. A total of 1125 men underwent randomization; the median follow-up was 34 months. There were 102 deaths in the enzalutamide group and 143 deaths in the standard-care group (hazard ratio, 0.67; 95% confidence interval [CI], 0.52 to 0.86; P = 0.002). Kaplan-Meier estimates of overall survival at 3 years were 80% (based on 94 events) in the enzalutamide group and 72% (based on 130 events) in the standard-care group. Better results with enzalutamide were also seen in PSA progression-free survival (174 and 333 events, respectively; hazard ratio, 0.39; P<0.001) and in clinical progression-free survival (167 and 320 events, respectively; hazard ratio, 0.40; P<0.001). Treatment discontinuation due to adverse events was more frequent in the enzalutamide group than in the standard-care group (33 events and 14 events, respectively). Fatigue was more common in the enzalutamide group; seizures occurred in 7 patients in the enzalutamide group (1%) and in no patients in the standard-care group. Enzalutamide was associated with significantly longer progression-free and overall survival than standard care in men with metastatic, hormone-sensitive prostate cancer receiving testosterone suppression. The enzalutamide group had a higher incidence of seizures and other toxic effects, especially among those treated with early docetaxel. (Funded by Astellas Scientific and Medical Affairs and others; ENZAMET (ANZUP 1304) ANZCTR number, ACTRN12614000110684; ClinicalTrials.gov number, NCT02446405; and EU Clinical Trials Register number, 2014-003190-42.).


Genetic determinants of reovirus pathogenesis in a murine model of respiratory infection.
Many viruses invade mucosal surfaces to establish infection in the host. Some viruses are restricted to mucosal surfaces, whereas others disseminate to sites of secondary replication. Studies of strain-specific differences in reovirus mucosal infection and systemic dissemination have enhanced an understanding of viral determinants and molecular mechanisms that regulate viral pathogenesis. After peroral inoculation, reovirus strain type 1 Lang replicates to high titers in the intestine and spreads systemically, whereas strain type 3 Dearing (T3D) does not. These differences segregate with the viral S1 gene segment, which encodes attachment protein σ1 and nonstructural protein σ1s. In this study, we define genetic determinants that regulate reovirus-induced pathology following intranasal inoculation and respiratory infection. We report that two laboratory isolates of T3D, T3D(C) and T3D(F), differ in the capacity to replicate in the respiratory tract and spread systemically; the T3D(C) isolate replicates to higher titers in the lungs and disseminates, while T3D(F) does not. Two nucleotide polymorphisms in the S1 gene influence these differences, and both S1 gene products are involved. T3D(C) amino acid polymorphisms in the tail and head domains of σ1 protein influence the sensitivity of virions to protease-mediated loss of infectivity. The T3D(C) polymorphism at nucleotide 77, which leads to coding changes in both S1 gene products, promotes systemic dissemination from the respiratory tract. A σ1s-null virus produces lower titers in the lung after intranasal inoculation and disseminates less efficiently to sites of secondary replication. These findings provide new insights into mechanisms underlying reovirus replication in the respiratory tract and systemic spread from the lung.


Sex Differences in Pharmacotherapy and Long-Term Outcomes in Patients With Ischaemic Heart Disease and Comorbid Left Ventricular Dysfunction.
Left ventricular (LV) dysfunction and ischaemic heart disease (IHD) are common among women. However, women tend to present later and are less likely to receive guideline-directed medical therapy (GDMT) compared with men. We analysed prospectively collected data (2005-2018) from a multicentre registry on GDMT 30 days after percutaneous coronary intervention in 13,015 patients with LV ejection fraction <50%. Guideline-directed medical therapy was defined as beta blocker, angiotensin-converting enzyme inhibitor/angiotensin receptor blocker±mineralocorticoid receptor antagonist. Long-term mortality was determined by linkage with the Australian National Death Index. Women represented 20% (2,634) of the total cohort. Mean age was 65±12 years. Women were on average >5 years, with higher body mass index and higher rates of hypertension, diabetes, renal dysfunction, prior stroke, and rheumatoid arthritis. Guideline-directed medical therapy was similar between sexes (73% vs 72%; p=0.58), although women were less likely to be on an angiotensin-converting enzyme inhibitor/angiotensin receptor blocker (80% vs 82%; p=0.02). Women were less likely to be on statin therapy (p<0.001) or a second antiplatelet agent (p=0.007). Women had higher unadjusted long-term mortality (25% vs 19%; p<0.001); however, there were no differences in long-term mortality between sexes on adjusted analysis (hazard ratio 0.99; 95% confidence interval 0.87-1.14; p=0.94). Rates of GDMT for LV dysfunction were high and similar between sexes; however, women were less likely to be on appropriate IHD secondary prevention. The increased unadjusted long-term mortality in women was attenuated in adjusted analysis, which highlights the need for optimisation of baseline risk to improve long-term outcomes of women with IHD and comorbid LV dysfunction.


Alpha1-antitrypsin inhibits the activity of the matriptase catalytic domain in vitro.
Matriptase is a type II transmembrane protease that is characterized by an N-terminal transmembrane and multiple extracellular domains, in addition to the conserved extracellular serine protease catalytic domain. The expression pattern of matriptase suggests that this protease may play broad roles in the biology of surface lining epithelial cells. In this study we report that alpha1-antitrypsin (AAT), an endogenous inhibitor of serine proteases, inhibits the catalytic domain of human recombinant matriptase in vitro. Co-incubation of AAT with matriptase (at a molar ratio 1:2) resulted in the formation of heat stable complexes, clearly seen in sodium dodecyl sulfate electrophoresis and Western blots. AAT was found to be a slow, tight-binding inhibitor of the catalytic domain of matriptase with a second order reaction rate constant of 0.31 x 10(3) M(-1)s(-1). Notably, the oxidized form of AAT, which lacks serine protease inhibitor activity, failed to generate matriptase complexes and to inhibit matriptase activity. Since matriptase is involved in a number of physiologic processes, including activation of epithelial sodium channels, our findings offer considerable new insights into new regulatory function of AAT in vivo.


An analysis of the sequences of the variable regions of Bence Jones proteins and myeloma light chains and their implications for antibody complementarity.
In an attempt to account for antibody specificity and complementarity in terms of structure, human kappa-, human lambda-, and mouse kappa-Bence Jones proteins and light chains are considered as a single population and the variable and constant regions are compared using the sequence data available. Statistical criteria are used in evaluating each position in the sequence as to whether it is essentially invariant or group-specific, subgroup-specific, species-specific, etc. Examination of the invariant residues of the variable and constant regions confirms the existence of a large number of invariant glycines, no invariant valine, lysine, and histidine, and only one invariant leucine and alanine in the variable region, as compared with the absence of invariant glycines and presence of three each of invariant alanine, leucine, and valine and two each of invariant lysine and histidine in the constant region. The unique role of glycine in the variable region is emphasized. Hydrophobicity of the invariant residues of the two regions is also evaluated. A parameter termed variability is defined and plotted against the position for the 107 residues of the variable region. Three stretches of unusually high variability are noted at residues 24-34, 50-56, and 89-97; variations in length have been found in the first and third of these. It is hypothesized that positions 24-34 and 89-97 contain the complementarity-determining residues of the light chain-those which make contact with the antigenic determinant. The heavy chain also has been reported to have a similar region of very high variability which would also participate in forming the antibody-combining site. It is postulated that the information for site complementarity is contained in some extrachromosomal DNA such as an episome and is incorporated by insertion into the DNA of the structural genes for the variable region of short linear sequences of nucleotides. The advantages and disadvantages of this hypothesis are discussed.


Adaptable and Automated Small UAV Deployments via Virtualization.
In this paper, we present a practical solution to support the adaptable and automated deployment of applications of Small Unmanned Aerial Vehicles (SUAVs). Our solution is based on virtualization technologies, and considers SUAVs as programmable network platforms capable of executing virtual functions and services, which may be dynamically selected according to the requirements specified by the operator of the aerial vehicles. This way, SUAVs can be flexibly and rapidly adapted to different missions with heterogeneous objectives. The design of our solution is based on Network Function Virtualization (NFV) technologies, developed under the umbrella of the fifth generation of mobile networks (5G), as well as on existing Internet protocol standards, including flying ad hoc network routing protocols. We implemented a functional prototype of our solution using well-known open source technologies, and we demonstrated its practical feasibility with the execution of an IP telephony service. This service was implemented as a set of virtualized network functions, which were automatically deployed and interconnected over an infrastructure of SUAVs, being the telephony service tested with real voice-over-IP terminals.


[Stage I or II Hodgkin's disease: more chemotherapy and less irradiation].
In Hodgkin's disease stage I or II, excellent survival rates have been reached by radiotherapy alone as well as by the combination of radiotherapy and chemotherapy. Unfortunately, patients surviving for longer than 10 to 15 years are at risk for late complications due to the irradiation, such as myocardial infarction and lung and breast cancer. The current treatment strategy is aimed at minimising these late effects without jeopardizing the good survival rates. Randomized trials, such as the Hodgkin trials of the European Organization for Research and Treatment of Cancer (EORTC), are the only way to ultimately determine the relative values of radiotherapy and chemotherapy. Every Hodgkin patient deserves being included in such a trial, because the survival results are optimal. In future clinical trials, radiotherapy alone will probably be replaced by chemotherapy combined with radiotherapy as the principal treatment modality.


Actual hazard of methyl bromide fumigation in soil disinfection.
Methyl bromide, a highly toxic and ready penetrating fumigant, is widely used against rodents, insects, mites, and a range of pathogenic organisms in soil, compost, and timber. To disinfect soil in greenhouses, methyl bromide is brought under pressure from outside by a vaporiser and blown on to ground under a polyethylene cover. The gas being three times heavier than air easily penetrates the ground. Depending on the local ventilation, a considerable amount of gas evaporates into the surrounding atmosphere, this emission being especially serious during the fumigation procedure and at the removal of the plastic cover. Previously, mechanical injection of methyl bromide on to the ground within closed areas was prohibited, since this technique exposed at least four disinfection workers at a time, who were provided with only a canister respirator, to gas concentrations of over 1000 ppm CH3Br. The present study established that fumigation with methyl bromide also carries risks for the well-protected worker inside, as well as for the one controlling the vaporiser. The concentration during application varies from 30 to 3000 ppm. Concentration in the air declines with time to 4 ppm CH3Br five days after application. Discarding the plastic sheet involves exposure to peak values as high as 200 ppm for a few seconds. On the ninth day after application, milling the soil can expose workers to up to 15 ppm; on the eleventh day no CH3Br concentration in the air could be found.


[Chemotherapy via tunneled central venous catheter left in place between drug courses. Prospective study in 98 ambulatory patients].
Chemotherapy is normally given during several months and requires a good vascular access. Repeated injections of cytotoxic agents are harmful for peripheral veins. We report here a prospective study on 98 ambulatory patients provided with a tunneled central venous catheter left in situ between drug administration. The catheter is inserted either in the subclavian or the internal jugular vein. Between chemotherapy drugs injection, heparin is given once a week in the catheter. Among the several advantages reported with this technique: easiness of drug injection, comfort of patients, possibility of new insertions in an other site in case of occlusion and the low cost. This technique is a "routine" at the Institut Curie since 5 years. Advantages and side effects of this technique are compared with arterio-venous shunting or totally implantable devices used by other authors.


Predicting miRNA Association with Corresponding Target Genes and Single Nucleotide Polymorphisms in Altered Renal Pathophysiology.
Translational research on miRNAs develops reliable biomarkers for diagnosis and prognosis of renal diseases. Bioinformatic analyses and systems biology could drive the research for knowing new informative miRNA targets. This study proposes an approach to identify miRNA specific significant target genes, and single nucleotide polymorphisms (SNPs) associated with renal pathophysiology. miRNAs were selected after removing duplicity, on the basis of techniques used, and disease spectrum width score. Target genes were predicted from different databases like miRWalk, miRTarBase, and DIANA-TarBase. SNPs were prioritized on the basis of target score and conserved energy score available in MirSNP database. miRNAs were characterized as "specific", "strong", "likely", "unlikely", and "irrelevant" biomarkers. PCR-SSP based genotyping was carried out to access the molecular profiling of hsa-miR-192 and TGF-β1 followed by quantitative real time PCR to analyze expression level of TGF-β1. The relative expression levels of mRNAs were analyzed by 2-ΔΔCt method. 170 renal associated miRNAs were found to be up-regulated, down-regulated or differentially expressed. Noticeably hsa-miR-192-3p expression was reported in nine diseases. 117 genes were associated with basic kidney diseases and end stage renal disease (ESRD). Threshold > 80% for 93 target genes was observed from mirSVR. Mutant genotypes for hsa-miR-192 (OR=4.64, p-value ≤ 0.0001) and its corresponding target gene TGF-β1 (OR = 0.70, p-value = 0.0351) showed susceptible association with ESRD. More so, patients possessing mutant allele of TGF-β1 showed elevated mRNA expression (Fold change = 9.83). Study proposed a new approach to identify specific miRNA biomarkers for particular diseases with corresponding target genes and SNPs and also highlighted the importance of hsa-miR-192 in renal diseases.


Beta-agonists and asthma: too much of a good thing?
In an unusual paradox, asthmatics who are chronically treated with bronchodilating beta-agonists sometimes experience a worsening of their condition. A new study describes one possible mechanism and reveals a potential new therapeutic target in the treatment of asthma.


Membrane environment modulates the pKa values of transmembrane helices.
In this work, we apply the recently developed constant pH molecular dynamics technique to study protonation equilibria of titratable side chains in the context of simple transmembrane (TM) helices and explore the effect of pH on their configurations in membrane bilayers. We observe that, despite a significant shift toward neutral states, considerable population of different side chains stay in the charged state that give rise to pKa values around 9.6 for Asp and Glu and 4.5 to 6 for His and Lys side chains, respectively. These charged states are highly stabilized by favorable interactions between head groups, water molecules, and the charged side chains that are facilitated by substantial changes in the configuration of the peptides. The pH dependent configurations and the measured pKa values are in good agreement with relatively recent solid state NMR measurements. Our results presented here demonstrate that all-atom constant pH molecular dynamics can be applied to membrane proteins and peptides to obtain reliable pKa values and pH dependent behavior for these systems.


Neuroprotective effect of memantine on hippocampal neurons in infantile rat hydrocephalus.
The effect of memantine administration on hippocampal neurons of the infantile rats with kaolin induced hydrocephalus was investigated. Hydrocephalus was induced by injecting kaolin into the cisterna magna of 3-weeks old Sprague-Dawley rat pups. One group received a single daily dose of 20mg/kg memantine i.p. following hydrocephalus induction for a period of two weeks. By the end of the two-week period, animals were radiologically evaluated by magnetic resonance imaging and then sacrificed to get their cerebrums removed. Both immunohistochemical analysis of nitric oxide synthase activity and quantification of spared neurons in CA1, CA2 and CA3 regions of hippocampus were performed. In hydrocephalus-induced rats considerable neuronal loss associated with significantly increased nitric oxide synthase immunoreactivity were determined in all hippocampal regions. However, memantine treated rats showed significantly higher number of spared neuron counts and reduced nitric oxide synthase immunoreactivity in CA1 and CA2 regions compared with the non-treated rats. The findings of the study show that hippocampal neurons may constitute important targets for injury secondary to hydrocephalic process in experimental infantile hydrocephalus. Early anti-excitotoxic treatment with memantine seems to have a neuroprotective effect especially in the CA1 and CA2 subunits of the hippocampus.


Tricyclic antidepressants for management of residual symptoms in inflammatory bowel disease.
Tricyclic antidepressants (TCAs) have efficacy in treating irritable bowel syndrome (IBS). Some clinicians use TCAs to treat residual symptoms in inflammatory bowel disease (IBD) patients already on decisive IBD therapy or with quiescent inflammation, although this strategy has not been formally studied. The aim of this study was to examine the efficacy of TCA therapy in IBD patients with residual symptoms, despite controlled inflammation, in a retrospective cohort study. Inclusion required initiation of TCA for persistent gastrointestinal symptoms. IBD patients had inactive or mildly active disease with persistent symptoms despite adequate IBD therapy as determined by their physician. Symptom response was compared with IBS patients. Established Likert scales were used to score baseline symptom severity (0=no symptoms, 3=severe symptoms) and TCA response (0=no improvement; 3=complete satisfaction). Eighty-one IBD [41.3±1.7 y, 56F; 58 Crohn's disease/23 ulcerative colitis (UC)] and 77 IBS (46.2±1.7 y, 60F) patients were initiated on a TCA therapy. Baseline symptom scores (IBD, 2.06±0.03; IBS, 2.12±0.04; P=0.15) and symptom response to TCA therapy (IBD, 1.46±0.09; IBS, 1.30±0.09; P=0.2) were similar in both the groups. At least moderate improvement (Likert score ≥2) on TCA was achieved by comparable proportions of patients (59.3% IBD vs. 46% IBS; P=0.09). Within IBD, response was better with UC than Crohn's disease (1.86±0.13 vs. 1.26±0.11, respectively, P=0.003). In a clinical practice setting, TCA use led to moderate improvement of residual gastrointestinal symptoms in IBD patients for whom escalation of IBD therapy was not planned. UC patients demonstrated higher therapeutic success. IBD symptom responses were similar to IBS patients.


Serum antibody response to lytic and latent Epstein-Barr virus antigens in undifferentiated nasopharyngeal carcinoma patients from an area of nonendemicity.
Epstein-Barr virus (EBV)-associated undifferentiated carcinoma of the nasopharyngeal type (UCNT) is highly prevalent in southeast China, where immunoglobulin A (IgA) antibodies to viral capsid antigen and early antigen (EA) represent important markers, routinely used to assist in diagnosing this malignancy. Our study aimed at determining the EBV serological profiles of 78 UCNT patients from Italy, an area of nonendemicity for this tumor, using different assays specific for both lytic and latent EBV antigens. Serum IgA against both EA and EBNA1 and IgG and IgA to the latent membrane protein 1 (LMP1), to EA, and to the EBV transactivator ZEBRA protein were assessed. These serological responses were then evaluated according to the clinicopathologic parameters at diagnosis. The sensitivities of the IgG assays were 37.7% for LMP1, 73.6% for EA, and 61.0% for ZEBRA. EA/EBNA1 IgA reactivity was 84.4%, and a high association (odds ratio [OR], 2.6; 95% confidence interval [CI], 1.7 to 4.0) with UCNT was observed. When EBV serological reactivities were analyzed according to the tumor, node, and metastasis staging system (TNM), a statistically significant association was found between N stage and IgG antibody rates for EA (OR, 3.6; 95% CI, 1.2 to 10.9) and ZEBRA (OR, 2.6; 95% CI, 1.2 to 5.5) and between M stage and IgG antibody rates for ZEBRA (OR, 7.1; 95% CI, 3.2 to 16.0) and LMP1 (OR, 14.0; 95% CI, 1.8 to 110.9). Our results show that no single serological marker allows the detection of all UCNT cases. EA/EBNA1 IgA represents a reliable marker for diagnosis, with a high predictive value also in areas where UCNT is not endemic, such as Italy. The analysis of serological results according to TNM classification is consistent with a progressive impairment of humoral immune response to EBV as the disease advances and may be used to improve the accuracy of diagnosis.


First-principles description of electrocatalytic characteristics of graphene-like materials.
Graphene-like materials (GLMs) have received much attention as a potential alternative to precious metal-based electrocatalysts. However, the description of their electrocatalytic characteristics may still need to be improved, especially under constant chemical potential. Unlike the case of conventional metal electrodes, the potential drop across the electrical double layer (ϕD) at the electrode-electrolyte interface can deviate substantially from the applied voltage (ϕapp) due to a shift of the Dirac point (eϕG) with charging. This may in turn significantly alter the interfacial capacitance (CT) and the relationship between ϕapp and free-energy change (ΔF). Hence, accurate evaluation of the electrode contribution is necessary to better understand and optimize the electrocatalytic properties of GLMs. In this work, we revisit and compare first-principles methods available to describe the ϕapp-∆F relation. Grand-canonical density functional theory is used to determine ΔF as a function of ϕapp or electrode potential (ϕq), from which the relative contribution of eϕG is estimated. In parallel, eϕG is directly extracted from a density functional theory analysis of the electronic structure of uncharged GLMs. The results of both methods are found to be in close agreement for pristine graphene, but their predictions deviate noticeably in the presence of adsorbates; the origin of the discrepancy is analyzed and explained. We then evaluate the application of the first-principle methods to prediction of the electrocatalytic processes, taking the reduction (hydrogenation) and oxidation (hydroxylation) reactions on pristine graphene as examples. Our work highlights the vital role of the modification of the electrode electronic structure in determining the electrocatalytic performance of GLMs.


Eat better & move more: a community-based program designed to improve diets and increase physical activity among older Americans.
We assessed outcomes of an integrated nutrition and exercise program designed for Older Americans Act Nutrition Program participants as part of the Administration on Aging's You Can! campaign. A 10-site intervention study was conducted. Preintervention and postintervention assessments focused on nutrition and physical activity stages of change, self-reported health status, dietary intakes, physical activity, and program satisfaction. Of 999 enrollees, the 620 who completed the program were aged 74.6 years on average; 82% were women, and 41% were members of racial/ethnic minority groups. Factors associated with program completion were site, health conditions, and nutrition risk. Seventy-three percent and 75% of participants, respectively, made a significant advance of 1 or more nutrition and physical activity stages of change; 24% reported improved health status. Daily intake of fruit increased 1 or more servings among 31% of participants; vegetables, 37%; and fiber, 33%. Daily steps increased 35%; blocks walked, 45%; and stairs climbed, 24%. Program satisfaction was 99%. This easy-to-implement program improves diets and activity levels. Local providers should offer more such programs with the goal of enabling older Americans to take simple steps toward successful aging.


What is the optimum minimum segment size used in step and shoot IMRT for prostate cancer?
Although the use of small segments in step and shoot IMRT provides better dose distribution, extremely small segments decrease treatment accuracy. The purpose of this study was to determine the optimum minimum segment size (MSS) in two-step optimization in prostate step and shoot IMRT with regard to both planning quality and dosimetric accuracy. The XiO treatment planning system and Oncor Impression Plus were used. Results showed that the difference in homogeneity index (HI), defined as the ratio of maximum to minimum doses for planning target volume, between the MSS 1.0 cm and 1.5 cm plans, and 2.0 cm plans, was 0.1%, and 9.6%, respectively. With regard to V107 of PTV, the volume receiving 107% of the prescribed dose of the PTV, the difference between MSS 1.0 cm and 1.5 cm was 2%. However, the value of the MSS 2.0 cm or greater plans was more than 2.5-fold that of the MSS 1.0 cm plan. With regard to maximum rectal dose, a significant difference was seen between the MSS 1.5 cm and 2.0 cm plans, whereas no significant difference was seen between the MSS 1.0 cm and 1.5 cm plans. Composite plan verification revealed a greater than 5% dose difference between planned and measured dose in many regions with the MSS 1.0 cm plan, but in only limited regions in the MSS 1.5 cm plan. Our data suggest that the MSS should be determined with regard to both planning quality and dosimetric accuracy.


Public and private perspectives on hospital conversions: proposed MUSC-Columbia/HCA "partnership".
Conversion has been defined as "any type of transaction that results in the shift of all or a substantial portion of the assets of nonprofit health care organizations to for-profit use" (Claxton and Colleagues, 1997:10-11). Not surprisingly, efforts at such conversion create political conflict and opposition within communities with some groups supporting and others opposing the intended conversion. This study looks at a particular effort at conversion: the proposed "partnership" between three hospitals of the Medical University of South Carolina (MUSC) and Columbia/HCA, the largest for-profit health services network in the U.S. Beginning in 1995, the three-year effort (now at an impasse) has drawn local and state policy-markers into increasing controversy and acrimony. Beyond the details, the episode also raises fundamental issues about the nature of public health care and the extent to which we can convert public health facilities to private control while preserving their essential public missions.


[Biological characteristics of coagulase-negative staphylococci isolated from conjunctivitis].
The biological properties of 101 strains of coagulase-negative staphylococci isolated from conjunctival secretions of conjunctivitis patients, as well as from healthy persons were studied. More than a half of these strains (66%) were identified as Staph. epidermidis. 18% of the strains differed from this species only by the absence of phosphatase activity. The existence of the phosphatase-negative variants of Staph. epidermidis was inferred from this fact. About 1/3 of the cultures could not be identified in accordance with the scheme of the International Subcommittee on the Taxonomy of Staphylococci and Micrococci. The biological activity of the cultures isolated from conjunctivitis patients did not differ from that of the cultures isolated from healthy persons. The frequent occurrence of hemolytic activity in the strains isolated from both conjunctivitis patients and healthy persons was noted (88% and 83%, respectively). The conclusion on the variegated character and lability of the phenotypic characteristics of staphylococci has been made.


The Role of the Transcription Factor E2F1 in Hepatocellular Carcinoma.
Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and the third leading cause of cancer-related death. Because of the fast growth, early hepatic metastasis and the multidrug resistance, the five-year survival rate is very low. Thus, the understanding of its biology can significantly contribute in identifying valuable targets for novel therapeutic approaches. In this regard, E2F1 may represent an interesting candidate. E2F1 is a transcription factor implicated in the regulation of many cellular processes including cell proliferation and apoptosis. Whereas the involvement of E2F1 in HCC has been recognized, its ability to act as a proliferative and/or apoptotic factor in HCC has not yet been clarified and, in this regard, an active debate is ongoing. The definition of E2F1 role in HCC is not a trivial aspect as it can have significant consequences for the development of novel therapeutic options with E2F1 as target. In this review, we present data about the reported proliferative/apoptotic effects as well as the dual (combined proliferation and apoptosis) functions of E2F1 in HCC discussing the molecular basis for this behavior. The data available so far indicate that the proliferative and apoptotic functions of E2F1 in HCC may coexist but the proliferative effect seems to be more pronounced than the apoptotic one.


The effect of numbers and locations of retentive holes placed on master casts on reducing the polymerization distortion of the maxillary complete denture.
Polymerization shrinkage is one of the disadvantages ofpolymethyl methacrylate resulting in distortion of resin denture bases. The aim of this study is to analyze the effect of numbers and locations of retentive holes on reducing polymerization distortion. Forty eight master casts were assigned as: control group (A); one anchoring hole (B); three anchoring holes (C); five anchoring holes (D).The gap distances between the denture bases and casts were measured. The data were analyzed with one-way analysis of variance (ANOVA) followed by the Post-Hoc LSD test. Mechanical anchorage reduced the gap distances in lateral and mid-palate significantly; and this decrease was affected by numbers and locations of anchoring holes significantly.


[Nocturnal enuresis in children--how to diagnose, how to treat?].
The aim of this article is to review International Children's Continence Society guidelines on the recommended diagnostic evaluation and therapy for children with nocturnal enuresis. Nocturnal enuresis (NE) is the condition describing the symptom of wetting during sleep above the age of 5 years. NE is one of the most common disorders among children. Enuresis is characterised as monosymptomatic nocturnal enuresis (MNE) if there are no additional voiding problems. Children with other daytime symptoms (daytime incontinence, urgency, frequency) and nocturnal enuresis are said to have non-monosymptomatic nocturnal enuresis (NMNE). A careful medical history, including bladder diary, physical examination, urinalysis, an ultrasound of the urinary tract system will usually provide sufficient information for the physician to arrive at a diagnosis. Urodynamic, radiologic and endoscopic evaluation are not necessary in children with monosymptomatic nocturnal enuresis. Two first line treatment options of MNE are currently recommended: nonpharmacologic treatment and pharmacologic treatment (desmopressin). Nonpharmacologic treatment of enuresis includes motivational therapy, bladder-training exercises, fluid and food intake and enuresis alarm. Before using alarm treatment or desmopressin, simple therapeutic interventions should be considered. Children with nocturnal poliuria and normal bladder capacity will be more sensitive to desmopressin.


Value and limits of endorectal ultrasonography for preoperative staging of rectal carcinoma.
In rectal cancer, the depth of tumor infiltration and metastatic involvement of lymph nodes are important prognostic factors. Endosonography of the rectum, combining the advantages of both endoscopy and sonography, provides information not available from other imaging diagnostic techniques. From January 1989 to December 1997, 85 patients affected by rectal carcinoma were submitted to preoperative evaluation with endorectal ultrasonography. In 75 cases the results obtained with the endosonography were compared to the histology of the resected specimens. Overall accuracy in staging depth of infiltration was 90.7%. Overstaging occurred in 4% of patients, whereas understaging occurred in 5.3%. In staging lymph nodal involvement, overall accuracy was 76%, sensitivity was 69.8%, specificity was 84.4%, positive predictive value was 85.7%, and negative predictive value was 67.5%. Endorectal ultrasound is a safe and accurate diagnostic method for staging both tumor invasion and lymph node metastatic involvement, and for selecting an appropriate surgical strategy in patients affected by rectal cancer.


Nuclear export signal consensus sequences defined using a localization-based yeast selection system.
Proteins bearing nuclear export signals (NESs) are translocated to the cytoplasm from the nucleus mainly through the CRM1-dependent pathway. However, the NES consensus sequence remains poorly defined, and there are currently no high-throughput methods for identifying NESs. In this study, we report the development of an efficient yeast selection system for detecting nuclear export activity as well as several reliable NES consensus sequences identified using this method. Our selection system is based on the nuclear export-dependent rescue of Tys1p, an essential cytoplasmic protein that has been artificially localized to the nucleus in a haploid Delta tys1 knockout strain. A screen of a random peptide library revealed 101 distinct CRM1-dependent NESs, which were classified into six patterns according to the conserved hydrophobic spacing. By combining mutational analyses, we have defined new NES consensus sequences with more specific and redundant residues than the traditional consensus sequence, which are consistent with most experimentally confirmed NESs. These NES consensus sequences should help identify functional NESs, and our selection system can be used to identify other targeting signals or proteins imported to specific subcellular compartments.


Lasallia pustulata lichen as possible natural antigenotoxic, antioxidant, antimicrobial and anticancer agent.
The methanol extract of the lichen Lasallia pustulata was tested for genotoxic, antioxidant, antimicrobial and anticancer activities. We did this using a cytokinesis block micronucleus (MN) assay on peripheral blood lymphocytes, by measuring free radical and superoxide anion scavenging activity, reducing power, determining of total phenolic compounds and determining the total flavonoid content, measuring the minimal inhibitory concentration by the broth microdilution method against five species of bacteria and five species of fungi and by using the microculture tetrazolium test on FemX (human melanoma) and LS174 (human colon carcinoma) cell lines. As a result of this study, we found that the methanol extract of L. pustulata did not modify the frequency of the MN and nuclear division index in comparison to untreated cells (p > 0.05). These results revealed that the methanol extract had moderate free radical scavenging activity with IC50 values of 395.56 μg/mL. Moreover, the extract tested had effective reducing power and superoxide anion radical scavenging. The values of the minimum inhibitory concentration against the tested microorganisms ranged from 0.625 to 20 mg/mL. In addition, the extract tested had strong anticancer activity against both cell lines with IC50 values of 46.67 and 71.71 μg/mL.


In vitro induction of tumor-specific immunity. I. Parameters of activation and cytotoxic reactivity of mouse lymphoid cells immunized in vitro against syngeneic and allogeneic plasma cell tumors.
Induction of tumor-specific immunity in vitro was accomplished by cocultivation of cortisone-resistant murine thymocytes or spleen cells with irradiated syngeneic plasma cell tumors (PCT). The cytotoxic activity generated could be detected in a short-term (51)Cr-release assay. Optimal cytotoxic activity against PCT-associated transplantation antigens (TATA) was generated after 7 days in culture. Unlike cytotoxic responses to tumor allografts in which the cytotoxic activity was directed against allogeneic transplantation antigens, the cytotoxic activity obtained in the syngeneic tumor system was specific to the immunizing syngeneic PCT. Similar parameters of induction of cytotoxic responses in in vitro tumor allograft responses and in the syngeneic tumor system suggested that both reactions are cell-mediated cytotoxic immune responses. With regard to the magnitude of cytotoxic responses obtained, allogeneic transplantation antigens induced about a 30-fold higher cytotoxic immune response than plasma cell TATA. The results are consistent with the concept that in vitro tumor allograft responses and in vitro responses against TATA of PCT are similar in quality, but differ in the magnitude of the cytotoxic response provoked.


Identification of the herbicide Surflan and its active ingredient oryzalin, a dinitrosulfonamide, as xenoestrogens.
Numerous environmental contaminants have been identified as endocrine disruptors (EDs)--substances that alter endocrine homeostasis by interfering with the biological action, production, or pharmacokinetics of endogenous hormones. Xenoestrogens are those EDs whose biological activity is similar to endogenous estrogen. This report presents data that identified Surflan, a proprietary herbicide emulsion, and its active ingredient oryzalin as xenoestrogens. In vitro, Surflan and oryzalin activated an estrogen-inducible reporter gene, and oryzalin competitively displaced 17beta-estradiol from the estrogen receptor. In vivo, Surflan and oryzalin induced expression of estrogen-regulated high-molecular-weight choriogenin genes in medaka (Oryzias latipes). These results are consistent with the characteristics of previously identified xenoestrogens and indicate that Surflan and oryzalin have the potential to adversely affect numerous estrogen-regulated biological processes.


High-dose methylprednisolone sodium succinate as a single agent in relapsed childhood acute lymphoblastic leukaemia.
Twenty children received methylprednisolone (1 g/m2), daily for 5 to 8 days, as initial single agent therapy for relapsed acute lymphoblastic leukaemia. Bone marrow blasts were reduced to less than 5% in 2 and 5-10% in 3 of 12 patients with bone marrow relapses. In 3/9 with central nervous system relapses the cerebrospinal fluid (CSF) blasts completely cleared and were reduced in 4 others. In two patients with testicular relapses there was shrinkage of tumour and one patient with a navicular bone relapse became pain free. Toxicity was minimal. These results indicate high-dose methylprednisolone is an effective agent, particularly in the treatment of established central nervous system (CNS) disease and could contribute to early CNS directed therapy in acute lymphoblastic leukaemia.


Whole-Genome Sequence of Synthesized Allopolyploids in Cucumis Reveals Insights into the Genome Evolution of Allopolyploidization.
The importance of allopolyploidy in plant evolution has been widely recognized. The genetic changes triggered by allopolyploidy, however, are not yet fully understood due to inconsistent phenomena reported across diverse species. The construction of synthetic polyploids offers a controlled approach to systematically reveal genomic changes that occur during the process of polyploidy. This study reports the first fully sequenced synthetic allopolyploid constructed from a cross between Cucumis sativus and C. hystrix, with high-quality assembly. The two subgenomes are confidently partitioned and the C. sativus-originated subgenome predominates over the C. hystrix-originated subgenome, retaining more sequences and showing higher homeologous gene expression. Most of the genomic changes emerge immediately after interspecific hybridization. Analysis of a series of genome sequences from several generations (S0, S4-S13) of C. ×hytivus confirms that genomic changes occurred in the very first generations, subsequently slowing down as the process of diploidization is initiated. The duplicated genome of the allopolyploid with double genes from both parents broadens the genetic base of C. ×hytivus, resulting in enhanced phenotypic plasticity. This study provides novel insights into plant polyploid genome evolution and demonstrates a promising strategy for the development of a wide array of novel plant species and varieties through artificial polyploidization.


Primitive organization of cytosolic Ca(2+) signals in hepatocytes from the little skate Raja erinacea.
Cytosolic Ca(2+) (Ca(i)(2+)) signals begin as polarized, inositol 1, 4,5-trisphosphate (InsP3)-mediated Ca(i)(2+) waves in mammalian epithelia, and this signaling pattern directs secretion together with other cell functions. To investigate whether Ca(i)(2+) signaling is similarly organized in elasmobranch epithelia, we examined Ca(i)(2+) signaling patterns and InsP3 receptor (InsP3R) expression in hepatocytes isolated from the little skate, Raja erinacea. Ca(i)(2+) signaling was examined by confocal microscopy, InsP3R expression by immunoblot, and the subcellular distribution of InsP3Rs by immunochemistry. ATP induced a rapid increase in Ca(i)(2+) in skate hepatocytes, as it does in mammalian hepatocytes. Unlike in mammalian hepatocytes, however, the Ca(i)(2+) increase in skate hepatocytes began randomly throughout the cell rather than in the apical region. In cells loaded with heparin ATP-induced Ca(i)(2+) signals were inhibited, but de-N-sulfated heparin was not inhibitory, suggesting that the increases in Ca(i)(2+) were mediated by InsP3. Immunoblot analysis showed that the type I but not the types II or III InsP3R was expressed in skate liver. Confocal immunofluorescence revealed that the InsP3R was distributed throughout the hepatocyte, rather than concentrated apically as in mammalian epithelia. These findings demonstrate that ATP-induced Ca(i)(2+) signals are mediated by InsP3 in skate hepatocytes, as they are in mammalian hepatocytes. However, in skate hepatocytes Ca(i)(2+) signals begin at loci throughout the cell rather than as an organized apical-to-basal Ca(i)(2+) wave, which is probably because the InsP3R is distributed throughout these cells. This primitive organization of Ca(i)(2+) signaling may in part explain the observation that Ca(2+)-mediated events such as secretion occur much less efficiently in elasmobranchs than in mammals.


Comparative genome analysis of 15 clinical Shigella flexneri strains regarding virulence and antibiotic resistance.
Shigellosis is the major cause of dysentery globally. It is mainly attributed to two Shigella species, Shigella sonnei and Shigella flexneri, which leads to approximately 165 million infections and 1.1 million deaths each year. Rapid increase and widening of spectrum in antibiotics resistance make Shigella hard to be adequately controlled through existing prevention and treatment measures. It has also been observed that enhanced virulence and advent of antibiotic resistance (AR) could arise almost simultaneously. However, genetic linkages between the two factors are missing or largely ignored, which hinders experimental verification of the relationship. In this study, we sequenced 15 clinically isolated S. flexneri strains. Genome assembly, annotation and comparison were performed through routine pipelines. Differential resistant profiles of all 15 S. flexneri strains to nine antibiotics were experimentally verified. Virulence factors (VFs) belonging to 4 categories and 31 functional groups from the Virulence Factor Database (VFDB) were used to screen all Shigella translated CDSs. Distribution patterns of virulence factors were analysed by correlating with the profiles of bacterial antibiotics resistance. In addition, multi-resistant S. flexneri strains were compared with antibiotic-sensitive strains by focusing on the abundance or scarcity of specific groups of VFs. By doing these, a clear view of the relationships between virulence factors and antibiotics resistance in Shigella could be achieved, which not only provides a set of genetic evidence to support the interactions between VFs and AR but could also be used as a guidance for further verification of the relationships through manipulating specific groups of virulence factors.


Control of glioblastoma tumorigenesis by feed-forward cytokine signaling.
EGFRvIII-STAT3 signaling is important in glioblastoma pathogenesis. Here, we identified the cytokine receptor OSMR as a direct target gene of the transcription factor STAT3 in mouse astrocytes and human brain tumor stem cells (BTSCs). We found that OSMR functioned as an essential co-receptor for EGFRvIII. OSMR formed a physical complex with EGFRvIII, and depletion of OSMR impaired EGFRvIII-STAT3 signaling. Conversely, pharmacological inhibition of EGFRvIII phosphorylation inhibited the EGFRvIII-OSMR interaction and activation of STAT3. EGFRvIII-OSMR signaling in tumors operated constitutively, whereas EGFR-OSMR signaling in nontumor cells was synergistically activated by the ligands EGF and OSM. Finally, knockdown of OSMR strongly suppressed cell proliferation and tumor growth of mouse glioblastoma cells and human BTSC xenografts in mice, and prolonged the lifespan of these mice. Our findings identify OSMR as a critical regulator of glioblastoma tumor growth that orchestrates a feed-forward signaling mechanism with EGFRvIII and STAT3 to drive tumorigenesis.


Management of biliary complications in the liver transplant patient.
Biliary complications after liver transplantation remain common. Resources for livers are limited, and these individuals are often ill, making nonoperative treatment and management attractive options. The endoscopic route for evaluation (endoscopic retrograde cholangiopancreatography) remains preferable, due to its safety profile, as opposed to the percutaneous route (percutaneous transhepatic cholangiography with percutaneous transhepatic biliary drainage), though the endoscopic route may not be possible in patients with a Roux-en-Y reconstruction. The two most common early complications include leaks from the anastomosis or cystic duct stump (of the donor or native duct) and obstruction at the surgical anastomosis. Nonoperative treatment is often successful in early complications. Late complications presenting with leaks and obstruction are often more difficult to treat nonoperatively and frequently require surgical treatment or retransplantation, though both endoscopic and percutaneous methods can be useful in the management of these complications or as a bridge to definitive surgical therapy.


Amenable mortality by household income and living arrangements: a linked register-based study of Finnish men and women in 2000-2007.
Mortality amenable to healthcare interventions has increasingly been used as an indirect indicator of the effect of healthcare on health inequalities. Studies have consistently shown socioeconomic differences in amenable mortality, but evidence on the joint effects of multiple socioeconomic and demographic factors is limited. We examined whether income and living arrangements have an independent effect on amenable mortality taking into account other dimensions of social position. The longitudinal and yearly updated individual level data were derived from different administrative registers and obtained from Statistics Finland. The data set includes an 11% random sample of all individuals aged 25-74 years at the end of 1999 and an 80% oversample of deaths in the follow-up period between 2000 and 2007. We used Cox proportional hazard regression with appropriate weights. We found twofold to threefold differences in amenable mortality between the top and bottom income quintiles. These differences were found to be largely attenuated by economic activity and living arrangements. We also found differences in amenable mortality by living arrangements suggesting that those living alone, as well as lone parents and those cohabiting have higher amenable mortality. These differences were largely independent of our indicators of socioeconomic position and economic activity. While our results give indirect support to the hypothesis that income differences in amenable mortality may be at least partially due to barriers in access to care, the large independent effects of living arrangements on amenable mortality suggest that seeking care may also have an impact.


A numerical study of the acoustic radiation due to eddy current-cryostat interactions.
To investigate the acoustic radiation due to eddy current-cryostat interactions and perform a qualitative analysis on noise reduction methods. In order to evaluate the sound pressure level (SPL) of the eddy current induced warm bore wall vibration, a Finite Element (FE) model was created to simulate the noises from both the warm bore wall vibration and the gradient coil assembly. For the SPL reduction of the warm bore wall vibration, we first improved the active shielding of the gradient coil, thus reducing the eddy current on the warm bore wall. A damping treatment was then applied to the warm bore wall to control the acoustic radiation. Initial simulations show that the SPL of the warm bore wall is higher than that of the gradient assembly with typical design shielding ratios at many frequencies. Subsequent simulation results of eddy current control and damping treatment application show that the average SPL reduction of the warm bore wall can be as high as 9.6 dB, and even higher in some frequency bands. Combining eddy current control and suggested damping scheme, the noise level in a MRI system can be effectively reduced.


Highly efficient hydrogen evolution of platinum via tuning the interfacial dissolved-gas concentration.
A facile perfluorooctanesulfonate (PFOS)-modulation strategy was developed to precisely control the dissolved-gas concentration at the electrode/gas/electrolyte interface for enhanced HER. With PFOS modulation, lowered dissolved-hydrogen concentrations at the catalytic interface and sufficient exposure of the surface active area can be achieved. Accordingly, relative to pure Pt, PFOS-modulated Pt possesses a remarkable HER performance.


A double-labeling investigation of the afferent connectivity to cortical areas V1 and V2 of the macaque monkey.
The afferent connectivity of areas V1 and V2 was investigated using the fluorescent dyes fast blue and diamidino yellow. Simultaneous injection of each dye in retinotopically corresponding regions of these areas gave rise to two afferent populations of labeled neurons in subcortical and cortical structures which project to both areas. These two populations showed a variable degree of overlap in their spatial distribution. Neurons labeled by both dyes (double-labeled neurons) which, therefore, project to both areas, were found in substantial numbers in these overlap zones. When the injections were made in non-retinotopically corresponding regions in the two areas, both populations of labeled cells overlapped extensively in the cortex but not in subcortical structures, suggesting that the laws governing the topography of these two types of connections are different. In the cortex, the labeled neurons extended from the fundus of the lunate sulcus to the fundus of the superior temporal sulcus. A few labeled neurons were also found in the inferior temporal cortex and the parahippocampal gyrus. In all cortical regions, corticocortical neurons projecting to V1 and V2 were found in both supra- and infragranular layers, although double-labeled neurons were more numerous in infragranular layers. With increasing distance from V1 there was an increase in the proportion of neurons labeled in infragranular layers. The comparative strength of input to V1 and V2 was computed and was found to be higher to V2 in all cortical regions except the superior temporal sulcus which projected equally heavily to both areas. The superior temporal sulcus also stood out in that of all cortical regions it contained the highest proportion of double-labeled neurons. Single- and double-labeled neurons were found in a number of subcortical structures including the lateral geniculate nucleus, the inferior and lateral pulvinar, the intralaminar nuclei, the nucleus basalis of Meynert, and the amygdala. The pattern of labeling in the lateral pulvinar was in agreement with the suggestion that this structure has a complex topographical organization containing at least a dual representation of the visual field (Bender, D. B. (1981) J. Neurophysiol. 46: 672-693). In the pulvinar complex, densities of labeled neurons permitted evaluation of the strength of input to V1 and V2, the latter being the strongest. These results demonstrate that areas V1 and V2 share a vast amount of common input from the same cortical and subcortical structures and that a number of neurons project to both areas via branching axons.


Chemokine receptor CCR5 and CXCR4 expression in HIV-associated kidney disease.
The chemokine receptors CCR5 and CXCR4 have been identified as essential coreceptors for entry of HIV-1 strains into susceptible cells. Direct infection of renal parenchymal cells has been implicated in the pathogenesis of HIV-associated renal disease, although data are conflicting. The localization of CCR5 and CXCR4 in kidneys with HIV-associated renal disease is unknown. Formalin-fixed, paraffin-embedded renal biopsies from patients with HIV-associated nephropathy (HIVAN) (n = 13), HIV-associated immune complex glomerulonephritis (n = 3), HIV-associated thrombotic microangiopathy (n = 1), and HIV-negative patients with collapsing glomerulopathy (n = 8) were analyzed in this study. Cellular sites of expression of CCR5 and CXCR4 were identified by immunohistochemistry and by in situ hybridization. The presence of HIV-1 was detected by immunohistochemistry and by in situ hybridization. Expression of both chemokine receptors CCR5 and CXCR4 was undetectable in intrinsic glomerular, tubular, and renovascular cells in all analyzed cases. In the presence of tubulointerstitial inflammation, CCR5 and CXCR4 expression was localized to infiltrating mononuclear leukocytes. HIV-1 protein was undetectable by immunohistochemistry in all cases of HIV-associated renal disease. HIV-1 RNA was identified in one case of HIVAN but was restricted to infiltrating leukocytes. HIV-1 RNA was not detected in intrinsic renal cells in all analyzed cases. Identifying the cellular expression of HIV-coreceptors CCR5 and CXCR4 may help to clarify which tissues are permissive for direct HIV infection. These data do not support a role of productive HIV-1 infection of renal parenchymal cells in the pathogenesis of HIV-associated renal disease.


Cell-mediated immunity in patients on long-term haemodialysis.
Several parameters of cell-mediated immunity were assessed in thirty patients with chronic renal failure treated with long-term haemodialysis. Lymphopenia was uncommon, and only two patients showed diminished numbers of thymus-derived peripheral blood lymphocytes. Skin test energy to three antigens was documented in only one patient. No serious infections with intracellular organisms were noted in any of the patients. Phytohaemagglutinin (PHA) induced DNA synthesis in eleven patients was lower than that shown by normal subjects. This poor responsiveness to PHA was not corrected by haemodialysis, and in only two patients was the response improved by substituting normal human plasma for uraemic plasma in the lymphocyte cultures. Thus with the exception of PHA response, other parameters of cell-mediated immunity were intact in this group of chronic stable haemodialysis patients.


On the role of retrosplenial cortex in long-lasting memory storage.
The retrosplenial cortex (RSC) is involved in a range of cognitive functions. However, its precise involvement in memory processing is unknown. Pharmacological and behavioral experiments demonstrate that protein synthesis and c-Fos expression in the anterior part of RSC (aRSC) are necessary late after training to maintain for many days a fear-motivated memory. Long-lasting memory storage is regulated by D1/D5 dopamine receptors in aRSC and depends on the functional interplay between dorsal hippocampus and aRSC. These results suggest that the RSC recapitulates some of the molecular events that occur in the hippocampus to maintain memory trace over time.


Haplotype association analysis of combining unrelated case-control and triads with consideration of population stratification.
Combining data when data are collected under different study designs, such as family trios and unrelated case-control samples, gains more power and is cost-effective than analyzing each data separately. However, a potential concern is population stratification (PS) among unrelated case-control samples and analyses integrating data should address this confounding effect. In this paper, we develop a simpler method, haplotype generalized linear model (HGLM), that tests and estimates haplotype effects on disease risk and allows for modification against PS for combining data. We proposed to combine information across aggregations of haplotype weighted-counts estimated from population case-control data and trio data separately, and to perform subsequent GLM analysis. Furthermore, we present a framework of analysis of variance based on haplotype weighted-counts for detecting whether it is appropriate to combine two data sources, as well as the modified HGLM with clustering methods for addressing PS. We evaluate the statistical properties in terms of the accuracy, false positive rate (FPR) and empirical power using simulated data with regard to various disease risks, sample sizes, multi-SNP haplotypes and the presence of PS. Our simulation results indicate that HGLM performs comparably well with the likelihood-based haplotype association analysis, particularly when the haplotype effects are moderate, but may not perform well when dealing with lengthy haplotypes for small sample sizes. In the presence of PS, the modified HGLM remains valid and has satisfactory nominal level and small bias. Overall, HGLM appears to be successful in combining data and is simple to implement in standard statistical software.


Exploring the application value of the modified Delphi method in the development process of acupuncture and moxibustion guideline recommendations based on the WFAS Clinical Practice Guideline for Female Urinary Incontinence.
Taking the recommendations development of the World Federation of Acupuncture-Moxibustion Societies (WFAS) standard Clinical Practice Guideline for Female Urinary Incontinence as an example, this study analyzed the consensus expert composition, specific consensus process, and results in the development of the guideline's recommendations. It systematically examined the advantages of using the modified Delphi method in the formation of recommendations for acupuncture and moxibustion clinical practice guideline, with the aim of providing reference for the development of acupuncture and moxibustion guidelines in the same field.


[Effect of proton irradiation of Tolosa-Hunt syndrome].
The authors have analysed a case of Tolosa-Hunt syndrome with a persistent recurring course and a gradual development of resistance to steroid therapy. Computerized tomography revealed a high density zone in the affected cavernous sinus of the patient. Differential diagnosis between a tumor and Tolosa-Hunt syndrome was made. The patient received 3 fractions of proton beam irradiation by the "piercing" method (the beam being 15 mm in diameter) at a dose of 60 Gy. After irradiation computerized tomography showed complete recovery of disturbed functions and disappearance of the focus. Four-year remission was observed. In our experience, it was the first case of the use of radiation therapy in Tolosa-Hunt syndrome.


[Image-guided radiotherapy: rational, modalities and results].
The objective of Image-Guided Radiotherapy (IGRT) is to take in account the inter- or/and intrafraction anatomic variations (organ motion and deformations) in order to improve treatment accuracy. The IGRT should therefore translate in a clinical benefit the recent advances in both tumor definition thanks to functional imaging, and dose distribution thanks to intensity modulated radiotherapy. The IGRT enables direct or indirect tumor visualization during radiation delivery. If the tumor position does not correspond with the theoretical location of target derived from planning system, the table is moved. In case of important uncertainties related to target deformation, a new planning can be discussed. IGRT is realized by different types of devices which can vary in principle and as well as in their implementation: from LINAC-integrated-kV (or MV)-Cone Beam CTs to helicoidal tomotherapy, Cyberknife and Novalis low-energy stereoscopic imaging system. These techniques led to a more rational choice of Planning Target Volume. Being recently introduced in practice, the clinical results of this technique are still limited. Nevertheless, until so far, IGRT has showed promising results with reports of minimal acute toxicity. This review describes IGRT for various tumor localizations. The dose delivered by on board imaging should be taken in account. A strong quality control is required for safety and proper prospective evaluation of the clinical benefit of IGRT.


Baculovirus Infection of Human Monocyte-Derived Dendritic Cells Restricts HIV-1 Replication.
Acquired immune deficiency syndrome (AIDS) is mainly caused by infection with human immunodeficiency virus-1 (HIV-1) and still poses a global threat for which we lack a protective or therapeutic vaccine. Dendritic cells (DCs) play a major role in the onset of HIV infection, providing one of the primary sites of HIV replication, and also act as viral reservoirs in vivo. Previous studies have shown that baculovirus (BV) induces strong host immune responses against infections and malignancies. In this study, we infected human monocyte-derived DCs with recombinant BV (AcCAG-gag) and showed that AcCAG-gag-infected human DCs underwent maturation and produced interferon alpha and other proinflammatory cytokines accompanied by increases in the mRNA and protein expression levels of APOBEC3 (A3A, A3F, and A3G), proteins associated with the inhibition of HIV-1 replication. Surprisingly, HIV-1 inhibition is also observed in human DCs infected with a wild-type BV, as determined by the production of inflammatory cytokines, the expression of A3, and a reduction in the p24 level. Our findings outline the mechanism underlying the inhibition of HIV-1 in BV-infected human DCs and pave the way for the use of BV as an effective tool for immunotherapy against HIV-1.


Reinforcement Enhancing Effects of Nicotine Via Patch and Nasal Spray.
Confirming preclinical findings, nicotine in humans (via smoking) enhances reinforcement from nondrug rewards. Recent demonstration of similar effects with nicotine via e-cigarettes suggests they may also occur when using nicotine replacement therapies (NRT). Effects of nicotine via NRT patch or nasal spray were assessed on responding reinforced by music, video, or monetary rewards, or for no reward (control). Nontreatment seeking smokers (N = 31) participated in three virtually identical experimental sessions, each following overnight abstinence (CO ≤ 10 ppm). In a fully within-subjects design using a double-dummy procedure, these sessions involved: (1) nicotine patch (Nicoderm 14 mg) plus placebo spray, (2) placebo patch plus nicotine spray (Nicotrol, 2 × 1 mg/trial), or (3) placebo patch plus placebo spray. Session order was counter-balanced. Relative to placebo, reinforced responding due to nicotine via spray or patch was greater for video reward (both p < .01) but not for music reward (both p > .10). Similar results for NRT spray and patch confirms preclinical findings indicating no difference between fast and slow nicotine delivery, respectively, on reinforcement enhancing effects. Withdrawal relief was unrelated to these effects of nicotine via NRT on nondrug reinforcement. Nicotine from NRT has some reinforcement enhancing effects in humans, possibly in a manner consistent with nicotine via e-cigarettes but not tobacco smoking. Our findings could suggest differential dose-dependency of available rewards to enhanced reinforcement by nicotine. Such effects may help contribute to the efficacy of NRT for aiding smoking cessation, but more research focusing on dose-dependency of these nicotine actions is needed. Acute nicotine from smoking enhances reinforced responding for nondrug sensory rewards. Yet, nonsmoked nicotine, including from NRT medications of patch and nasal spray, may act more selectively across rewards, perhaps due to lower dosing exposure. Our results suggest that nicotine via NRT enhances responding for visual (video) reward, but not from auditory (music) reward, just as in prior results using e-cigarettes. Withdrawal relief from NRT was unrelated to reinforced responding, consistent with positive (and not negative) reinforcement from this nicotine. Further research evaluating the dose-response effects of nicotine may clarify differences in enhanced reinforcement depending on the type of available reward.


Occult orthopaedic trauma in the multiply injured patient.
A retrospective review of 111 multitrauma patients revealed that of 401 orthopaedic injuries, 24 injuries (6%) were not initially diagnosed in 20 patients. Patients with occult injuries tended to have greater overall trauma, as reflected by lower trauma and lower Glasgow coma scores and longer hospital and intensive-care unit stays. Twenty prospectively identified cases were added to the series to further define risk factors. Seventy percent of occult bony injuries were ultimately diagnosed by physical examination and plain radiographs alone. Only 27% of cases required sophisticated imaging techniques for diagnosis. Based on these 44 cases of occult injuries in multitrauma victims, the following risk factors were identified: (1) significant multisystem trauma with another more apparent orthopaedic injury within the same extremity, (2) trauma victim too unstable for full initial orthopaedic evaluation, (3) altered sensorium, (4) hastily applied emergency splint obscuring a less apparent injury, (5) poor quality or inadequate initial radiographs, and (6) inadequate significance assigned to minor signs/symptoms in a major trauma victim. Due to the nature and extent of the overall trauma, all injuries cannot be diagnosed on initial patient evaluation.


Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD.
This paper aims to screen and identify sphere clone cells with characteristics similar to cancer stem cells in human gallbladder cancer cell line GBC-SD. GBC-SD cells were cultured in a serum-free culture medium with different concentrations of the chemotherapeutic drug cisplatin for generating sphere clones. The mRNA expressions of stem cell-related genes CD133, OCT-4, Nanog, and drug resistance genes ABCG2 and MDR-1 in sphere clones were detected by quantitative real-time polymerase chain reaction (PCR). Stem cell markers were also analyzed by flow cytometry and immunofluorescent staining. Different amounts of sphere clones were injected into nude mice to test their abilities to form tumors. Sphere clones were formed in serum-free culture medium containing cisplatin (30 μmol/L). Flow cytometry results demonstrated that the sphere clones expressed high levels of stem cell markers CD133(+) (97.6%) and CD44(+) (77.9%) and low levels of CD24(+) (2.3%). These clones also overexpressed the drug resistance genes ABCG2 and MDR-1. Quantitative real-time PCR showed that sphere clones expressed stem cell genes Nanog and OCT-4 284 and 266 times, respectively, more than those in the original GBC-SD cells. Immunofluorescent staining showed that sphere clones overexpressed OCT-4, Nanog, and SOX-2, and low expressed MUC1 and vimentin. Tumor formation experiments showed that 1×10(3) sphere clone cells could induce much larger tumors in nude mice than 1×10(5) GBC-SD cells. In conclusion, sphere clones of gallbladder cancer with stem cell-like characteristics can be obtained using suspension cultures of GBC-SD cells in serum-free culture medium containing cisplatin.


Both cleavage products of the mCLCA3 protein are secreted soluble proteins.
Members of the chloride channels, calcium-activated (CLCA) family of proteins and in particular the murine mCLCA3 (alias gob-5) and its human ortholog hCLCA1 have been identified as clinically relevant molecules in diseases with secretory dysfunctions including asthma and cystic fibrosis. Initial studies have indicated that these proteins evoke a calcium-activated chloride conductance when transfected into human embryonic kidney cells 293 cells. However, it is not yet clear whether the CLCA proteins form chloride channels per se or function as mediators of other, yet unknown chloride channels. Here, we present a systematic biochemical analysis of the posttranslational processing and intracellular trafficking of the mCLCA3 protein. Pulse-chase experiments after metabolic protein labeling of mCLCA3-transfected COS-1 or human embryonic kidney 293 cells revealed cleavage of a primary 110-kDa mCLCA3 translation product in the endoplasmic reticulum into a 75-kDa amino-terminal and a 35-kDa carboxyl-terminal protein that were glycosylated and remained physically associated with each other. Confocal fluorescent analyses identified both cleavage products in vesicles of the secretory pathway. Neither cleavage product was associated with the cell membrane at any time. Instead, both subunits were fully secreted into the extracellular environment as a soluble complex of two glycoproteins. These results suggest that the two mCLCA3 cleavage products cannot form an anion channel on their own but may instead act as extracellular signaling molecules. Furthermore, our results point toward significant structural differences between mCLCA3 and its human ortholog, hCLCA1, which is thought to be a single, non-integral membrane protein.