Genetics and dementia nosology.
In preparation for the development of the Diagnostic and Statistical Manual of Mental Disorders (5th ed), the American Psychiatric Association convened workshops reviewing scientific evidence relevant to diagnosis of dementia. One of the domains covered was genetics, which is reviewed here. The following areas are reviewed: genetic data on Alzheimer's disease and other dementias; the impact of nosology on genetic research in terms of its potential to improve diagnostic sensitivity and specificity and to decrease heterogeneity; the impact of genetic research findings on nosology, specific diagnostic criteria, and subtypes; and recommendations and future directions. The focus is on Alzheimer's disease, where more genetic data are available, and other dementias are reviewed more briefly. In addition, a separate section reviews the relationship of genetic findings and mild cognitive impairment, a boundary zone between normal aging and dementia, particularly Alzheimer's disease.


The protective effect of melatonin on ischemia-reperfusion injury in the groin (inferior epigastric) flap model in rats.
Inadequate blood perfusion and ischemia-reperfusion (I/R) injury in the surgical skin flap are believed to be the major factors that cause harmful changes within the tissue and vasculature, resulting in flap necrosis. Reactive oxygen radical species (ROS), in part, are believed to play an important role in this injury. Melatonin, in many physiological conditions, has been shown to have direct and indirect antioxidative effects and free-radical-scavenging properties. Therefore, it may have a beneficial effect on I/R-induced flap injury. In this study, the possible protective effects of melatonin were investigated in I/R injury of rat epigastric (axial pattern) flaps. Ischemia was achieved for 12 h by occlusion of inferior epigastric artery. Melatonin or vehicle was administered 1 h before flap elevation and was continued for 6 days after ischemia. I/R injury elevated malondialdehyde (MDA), an end product of lipid peroxidation, and nitric oxide (NO) levels while the glutathione (GSH) content was reduced. Myeloperoxidase (MPO) activity, which is known to be related to tissue neutrophil accumulation, was found to be statistically higher in the I/R group when compared with the sham group. Administration of melatonin significantly decreased MDA, NO and MPO levels and elevated the GSH content. Moreover, melatonin reduced the flap necrosis area, which was determined using a planimetric method. In conclusion, melatonin, a potent scavenger of free radicals, plays a major role in preventing the inferior epigastric arterial I/R-induced flap necrosis, based on planimetric flap survival and biochemical results. The beneficial effects of melatonin in I/R injury implies the involvement of free radicals in flap damage.


Enhancing genomics-based outbreak detection of endemic Salmonella enterica serovar Typhimurium using dynamic thresholds.
Salmonella enterica serovar Typhimurium is the leading cause of salmonellosis in Australia, and the ability to identify outbreaks and their sources is vital to public health. Here, we examined the utility of whole-genome sequencing (WGS), including complete genome sequencing with Oxford Nanopore technologies, in examining 105 isolates from an endemic multi-locus variable number tandem repeat analysis (MLVA) type over 5 years. The MLVA type was very homogeneous, with 90 % of the isolates falling into groups with a five SNP cut-off. We developed a new two-step approach for outbreak detection using WGS. The first clustering at a zero single nucleotide polymorphism (SNP) cut-off was used to detect outbreak clusters that each occurred within a 4 week window and then a second clustering with dynamically increased SNP cut-offs were used to generate outbreak investigation clusters capable of identifying all outbreak cases. This approach offered optimal specificity and sensitivity for outbreak detection and investigation, in particular of those caused by endemic MLVA types or clones with low genetic diversity. We further showed that inclusion of complete genome sequences detected no additional mutational events for genomic outbreak surveillance. Phylogenetic analysis found that the MLVA type was likely to have been derived recently from a single source that persisted over 5 years, and seeded numerous sporadic infections and outbreaks. Our findings suggest that SNP cut-offs for outbreak cluster detection and public-health surveillance should be based on the local diversity of the relevant strains over time. These findings have general applicability to outbreak detection of bacterial pathogens.


Prospective Evaluation of Zoledronic Acid in the Treatment of Bone Metastases from Differentiated Thyroid Carcinoma.
The objectives of this study were to prospectively delineate the efficacy and safety of zoledronic acid for treating bone metastases (BM) from differentiated thyroid carcinoma (DTC), and to evaluate the relationships between levels of bone metabolic markers and occurrence of skeletal-related events (SREs). This was a prospective, single-arm, single-center study. Nineteen patients with BM from DTC were assigned to receive zoledronic acid therapy every 4-5 weeks. Imaging studies for sites of BM were conducted every 6 months, and levels of bone metabolic markers, including serum bone-specific alkaline phosphatase (BAP) and urinary N-telopeptide of type I collagen (NTx), were assessed every 3 months. To evaluate the efficacy of zoledronic acid use, data of SREs were compared with those of 16 historical controls. SREs developed in eight patients (42 %), but metastatic spinal cord compression (MSCC) appeared in only one patient. Pain scores were ameliorated in five patients, but aggravated in six patients. Ten patients had stable disease, six showed progressive disease, and none showed partial or complete response during the observation period. Decreases in levels of bone metabolic markers were observed in ten patients for BAP and 15 for NTx. However, no significant correlations were identified between changes in bone metabolic marker levels and development of SREs. Zoledronic acid may offer a mainstay of multidisciplinary treatment for patients with BM for the purpose of reducing SREs. Levels of serum BAP and urinary NTx do not appear reliable as indicators of amelioration of BM symptoms.


Long-term outcome of pyogenic vertebral osteomyelitis: a cohort study of 260 patients.
The long-term outcome of patients with pyogenic vertebral osteomyelitis (PVO) has not been fully assessed. We conducted a retrospective cohort study to describe the long-term outcome of PVO and to assess risk factors for treatment failure in patients evaluated at our institution between 1994 and 2002. Patients were observed until July 1, 2013. Two hundred sixty patients with PVO were included in this study. Twenty-seven percent (70) of patients developed their infection after an invasive spinal procedure. Staphylococcus aureus accounted for 40% (103) of infections. Forty-nine percent (128) of patients underwent spinal surgery as part of their initial therapy. The median duration of parenteral antimicrobial therapy was 42 days (interquartile range, 38-53). The estimated 2-, 5-, and 10-year cumulative probability of treatment failure-free survival was 72%, 69%, and 69%, respectively. Seventy-five percent of patients who developed treatment failure did so within 4.7 months of diagnosis. Residual neurological defects and persistent back pain were seen in 16% and 32% of patients, respectively. In a multivariate analysis, longer duration of symptoms before diagnosis and having an infection with S. aureus were associated with increased risk of treatment failure. Increasing duration of symptoms and infection with S. aureus were associated with treatment failure in patients with PVO. Most treatment failures occurred early after initiation of treatment. Pyogenic vertebral osteomyelitis is associated with a high 2-year failure rate. Persistent neurological deficits and back pain are common after therapy.


Influence of lipid composition of model membranes on methacrylate antimicrobial polymer-membrane interactions.
Using atomistic molecular dynamics simulations, the role of lipid composition in the interactions of multiple methacrylate antimicrobial polymer agents with model membranes, and the consequent response of the membranes is studied. In our earlier study, methacrylate polymers were observed to induce phase demixing and associated thickness mismatch in a POPE-POPG model microbial membrane. In this work, we probe (1) the role of varying the degree of saturation in lipid acyl chains in the membrane interactions of methacrylate polymers, and (2) whether electrostatics (addition of anionic lipids) can influence the interactions of the polymers with model mammalian membranes. Lipid composition is observed to significantly modify membrane-polymer interactions, leading to differences in both the mode of partitioning and the conformations adopted by the polymers, in addition to impacting membrane properties differently. The results strongly suggest that the oft-cited electrostatic interactions between the antimicrobial agents and the microbial membranes do not fully account for the recognition and subsequent partitioning of the antimicrobial agents. The ability of the methacrylate polymers to sense interfacial lipid packing defects, determined by the PE/PC head groups of lipids, is also found to be influential in their membrane partitioning. Deliberate inclusion of charged anionic lipids into a model mammalian membrane, leading to additional favorable electrostatics, does not reproduce a similar polymer partitioning mechanism to that in its microbial counterpart. The differences observed in the interactions of methacrylate polymers with the various model membranes can be instrumental in extending our understanding of underlying modes of membrane disruption by general antimicrobial agents as well.


Therapeutic Anticoagulation with Heparin in Noncritically Ill Patients with Covid-19.
Thrombosis and inflammation may contribute to the risk of death and complications among patients with coronavirus disease 2019 (Covid-19). We hypothesized that therapeutic-dose anticoagulation may improve outcomes in noncritically ill patients who are hospitalized with Covid-19. In this open-label, adaptive, multiplatform, controlled trial, we randomly assigned patients who were hospitalized with Covid-19 and who were not critically ill (which was defined as an absence of critical care-level organ support at enrollment) to receive pragmatically defined regimens of either therapeutic-dose anticoagulation with heparin or usual-care pharmacologic thromboprophylaxis. The primary outcome was organ support-free days, evaluated on an ordinal scale that combined in-hospital death (assigned a value of -1) and the number of days free of cardiovascular or respiratory organ support up to day 21 among patients who survived to hospital discharge. This outcome was evaluated with the use of a Bayesian statistical model for all patients and according to the baseline d-dimer level. The trial was stopped when prespecified criteria for the superiority of therapeutic-dose anticoagulation were met. Among 2219 patients in the final analysis, the probability that therapeutic-dose anticoagulation increased organ support-free days as compared with usual-care thromboprophylaxis was 98.6% (adjusted odds ratio, 1.27; 95% credible interval, 1.03 to 1.58). The adjusted absolute between-group difference in survival until hospital discharge without organ support favoring therapeutic-dose anticoagulation was 4.0 percentage points (95% credible interval, 0.5 to 7.2). The final probability of the superiority of therapeutic-dose anticoagulation over usual-care thromboprophylaxis was 97.3% in the high d-dimer cohort, 92.9% in the low d-dimer cohort, and 97.3% in the unknown d-dimer cohort. Major bleeding occurred in 1.9% of the patients receiving therapeutic-dose anticoagulation and in 0.9% of those receiving thromboprophylaxis. In noncritically ill patients with Covid-19, an initial strategy of therapeutic-dose anticoagulation with heparin increased the probability of survival to hospital discharge with reduced use of cardiovascular or respiratory organ support as compared with usual-care thromboprophylaxis. (ATTACC, ACTIV-4a, and REMAP-CAP ClinicalTrials.gov numbers, NCT04372589, NCT04505774, NCT04359277, and NCT02735707.).


Genomic analysis of the nuclear receptor family: new insights into structure, regulation, and evolution from the rat genome.
Completion of the Rattus norvegicus genome sequence enabled a global inventory and analysis of the nuclear receptors (NRs) in three mammalian species. Forty-nine NR members were found in mouse, 48 in human. Forty-seven were found in the rat, with gaps at the locations expected for the other two. Pairwise comparisons of their distribution in rat, mouse, and human identified 11 syntenic NR gene blocks, including three small clusters of two or three closely related genes, each spanning 40 kb to 1700 kb. The exon structure of the ligand-binding domain suggests that exon shuffling has played a role in the evolution of this family. An invariant splice junction in all members of the NR family except LXRbeta suggests a functional role for the intron. The ligand-binding domains of PXR and CAR are among the most divergent in the family. Their higher nucleotide substitution rates may be related to the central role played by these two NRs in the metabolism of the foreign compounds and may have resulted from limited positive selection.


Impact of late incomplete stent apposition after sirolimus-eluting stent implantation on 4-year clinical events: intravascular ultrasound analysis from the multicentre, randomised, RAVEL, E-SIRIUS and SIRIUS trials.
The impact of incomplete stent apposition (ISA) after drug-eluting stent implantation determined by intravascular ultrasound (IVUS) on late clinical events is not well defined. To evaluate the clinical impact of ISA after sirolimus-eluting stent (SES) placement during a follow-up period of 4 years. Pooled analysis from the RAVEL, E-SIRIUS and SIRIUS trials, three randomised, multicentre studies comparing SES and bare-metal stents (BMS). IVUS at angiographic follow-up was available in 325 patients (SES: n = 180, BMS: n = 145). IVUS images were reviewed for the presence of ISA defined as one or more unapposed stent struts. Clinical follow-up was available for a 4-year period in all patients. Frequency, predictors and clinical sequel of ISA at follow-up after SES and BMS implantation were determined. ISA at follow-up was more common after SES (n = 45 (25%)) than after BMS (n = 12 (8.3%), p<0.001). Canadian Cardiology Society class III or IV angina at stent implantation (odds ratio (OR) = 4.69, 95% CI 2.15 to 10.23, p<0.001) and absence of diabetes (OR = 3.42, 95% CI 1.05 to 11.1, p = 0.041) were predictors of ISA at follow-up after SES placement. Rate of myocardial infarction tended to be slightly higher for ISA than for non-ISA patients. When SES patients only were considered, major adverse cardiac event free survival at 4 years was identical for those with and without ISA at follow-up (11.1% vs 16.3%, p = 0.48). ISA at follow-up is more common after SES implantation than after BMS implantation. Considering the current very sensitive IVUS definition, ISA appears to be an IVUS finding without significant impact on the incidence of major adverse cardiac events even during long-term follow-up.


Heterogeneous response to HAART across a diverse population of people living with HIV: results from the ANRS-EN12-VESPA Study.
Benefits from HAART may be heterogeneous across people living with HIV. We measured the differences in the rate of HAART failure across the various subgroups represented at the level of a country. We used data from a national representative sample of people living with HIV and followed at hospital in France (ANRS-EN12-VESPA Study). Analyses were restricted to 896 participants on HAART for 6 months or more, who were antiretroviral naive at HAART initiation. Different indicators of treatment failure were defined: immunological failure (absence of an increase of 100 CD4 cells/microl or more); immunovirological failure (CD4 cell count of 200 cells/microl or less and detectable HIV-RNA); clinical failure (occurrence of an AIDS-defining illness more than 3 months after HAART initiation). Differences in the frequency of treatment failure were measured using logistic regression models adjusted for major established determinants of response to HAART. Overall, 19.6% of the study participants experienced immunological failure, 3.4% immunovirological failure, and 3.0% clinical failure, with substantial variations across the various subgroups. Compared with homo/bisexual men, migrants had higher rates of immunological failure (adjusted odds ratio 2.27, 95% confidence interval 1.14-4.56 for migrant men and 2.19, 1.17-4.08 for migrant women), immunovirological failure (8.23, 1.77-38.33 and 6.91, 1.03-46.32), and clinical failure (4.60, 1.01-20.86 and 4.22, 0.84-21.17). In France, migrants consistently appear to be at increased risk of treatment failure compared with other people with HIV. Understanding the reasons underlying such heterogeneity in terms of living conditions and educational/cultural background will be important.


Kinetic evidence for a common transport route of benzylpenicillin and probenecid by freshly prepared hepatocytes in rats. Influence of sodium ion, organic anions, amino acids and peptides on benzylpenicillin uptake.
Hepatic transport system of benzylpenicillin was characterized by using freshly prepared rat hepatocytes. Uptake of benzylpenicillin and cefpiramide did not require the presence of sodium ion in the incubation medium. No influence of several kinds of amino acids (leucine, histidine, phenylalanine, valine, alanine, glutamic acid and glycine) and peptides (prolyl-leucine, glycyl-glycine, glycyl-sarcosine, glycyl-leucine and gamma-glutamyl-cysteinylglycine) was observed for benzylpenicillin uptake into hepatocytes. Taurocholic acid and cholic acid significantly inhibited benzylpenicillin uptake. The kinetic study revealed that taurocholic acid inhibited benzylpenicillin uptake in a noncompetitive fashion. A similar effect of benzylpenicillin on taurocholic acid uptake was observed, suggesting that the affinity site of the hepatic transport carrier of benzylpenicillin is distinct from that of taurocholic acid. It is noteworthy that p-aminohippuric acid and p-acetylaminohippuric acid did not inhibit benzylpenicillin uptake. In contrast to the mutual inhibition behavior of benzylpenicillin and taurocholic acid, benzylpenicillin is fully and competitively inhibited by the simultaneous addition of probenecid. The inhibition constant Ki value of probenecid was calculated to be 0.322 mM. The uptake of probenecid was also significantly inhibited by benzylpenicillin. It is postulated that the affinity site of benzylpenicillin transport carrier is the same as that of probenecid and that the whole process of the benzylpenicillin transport system is common, at least in part, to the probenecid transport process in the liver.


Leveraging state immunization information systems to measure the effectiveness of rotavirus vaccine.
Electronic immunization information systems (IISs) are now established in almost all US states. We used the IIS in Minnesota, Georgia, and Connecticut for immunization data and as the source of 1 of 2 control groups to measure pentavalent rotavirus vaccine (RV5) effectiveness (VE) using case-control methodology. Case-subjects were vaccine-eligible children who presented to 1 of 5 hospitals or emergency departments with gastroenteritis and had rotavirus antigen detected in stool during any of 3 rotavirus seasons (2007-2009). Two control groups were used: children with gastroenteritis who tested negative for rotavirus and children from the IIS matched by zip code and birth date. In Minnesota and Georgia, immunization records for rotavirus-positive and -negative children were also obtained from providers. Overall, 402 (96%) rotavirus case-subjects and 825 (97%) rotavirus-negative controls who met eligibility criteria were found in the IISs. Ten IIS controls were identified for each case. VE estimates for RV5 were similar across control groups, immunization data sources, and states. VE point estimates for 3 vs 0 doses were 89% to 94% among children aged 8 months or older and 86% to 92% among those aged 24 months or older. VE for 2 doses was ≥90% among children aged 8 months or older, and VE for 1 dose was 66% among those aged 6 weeks through 5 months. Three RV5 doses confer sustained protection against rotavirus disease during the first 3 years of life in US children. Two RV5 doses also seem to provide good protection. IISs can be valuable tools for assessing the effectiveness of vaccines administered to young children.


Development and characterization of various osteoarthritis models for tissue engineering.
Osteoarthritis (OA) is characterized by a progressive loss of articular cartilage, subchondral bone sclerosis and synovial inflammation and is the most common chronic condition worldwide today. However, most treatments have focused on pain relief and OA symptoms. For these reasons, many ongoing studies are currently trying to develop efficient and successful therapies based on its pathology. Animal models that mimic the histopathology and symptoms of OA have a critical role in OA research and make it possible to investigate both secondary osteoarthritic changes due to a precedent event such as traumatic injury and naturally occurring changes for the development of therapeutics which can be tested in preclinical and clinical OA trials. We induced OA in various animal models including rats, rabbits and guinea pigs by chemical, surgical and naturally occurring methods. In particular, the Dunkin-Hartley guinea pig is very attractive as an OA animal model because OA slowly progresses which is similar to human primary OA. Thus, this animal model mimics the pathophysiological process and environment of human primary OA. Besides the spontaneous OA model, anterior cruciate ligament transection (ACLT) with medial meniscectomy and bilateral ovariectomy (OVX) as well as a chemical technique using sodium monoiodoacetate (MIA) were used to induce OA. We found that ACLT in the rat model induced OA changes in the histology and micro-CT image compared to OVX. The osteoarthritic change significantly increased following ACLT surgery in the rabbit model. Furthermore, we identified that OA pathogenic changes occurred in a time-dependent manner in spontaneous Dunkin-Hartley guinea pigs. The MIA injection model is a rapid and minimally invasive method for inducing OA in animal models, whereas the spontaneous OA model has a slow and gradual progression of OA similar to human primary OA. We observed that histological OA change was extraordinarily increased at 9 ½ months in the spontaneous OA model, and thus, the grade was similar with that of the MIA model. Therefore, this study reports on OA pathology using various animal models as well as the spontaneous results naturally occurring in an OA animal model which had developed cartilage lesions and progressive osteoarthritic changes.


SU-E-J-145: Implementing 4D XCAT Phantom for 4D Radiotherapy Research.
To characterize and implement the 4D Integrated Extended Cardiac Torso (XCAT) digital phantom for 4D radiotherapy (RT) application. A computer program was developed to facilitate the characterization and implementation of the 4D XCAT phantom. The program can (1) generate 4D XCAT images with customized parameter files; (2) review 4D XCAT images; (3) generate composite images from 4D XCAT images; (4) track motion of selected region-of-interested (ROI); (5) convert XCAT raw binary images into DICOM format; (6) analyze clinically acquired 4DCT images and Real-time Position Management (RPM) respiratory signal. Validation of the motion tracking algorithm was made by comparing to manual method. Major characteristics of the 4D XCAT phantom were studied including the dependence of lesion motion on its location/size and inputted diaphragm profile. An end-to-end test from image generation to treatment planning was also performed. The comparison between motion tracking and manual measurements of lesion motion trajectory showed a small difference between the two (mean difference in motion amplitude: 1.2 mm). The maximum lesion motion decreased nearly linearly (R2=0.97) as its distance to the diaphragm (DD) increased. At any given DD, lesion motion amplitude increased nearly linearly (R2 range: 0.89 to 0.95) as the inputted diaphragm motion increased. For a given diaphragm motion, the lesion motion is independent f the lesion size at any given DD. The 4D XCAT phantom can closely reproduce irregular breathing profile: the mean difference in motion amplitude between the inputted and the measured motion profile was 1.4 mm. The end-to-end test showed that clinically comparable treatment plans can be generated successfully based on 4D XCAT images. An integrated computer program has been developed to generate, review, analyze, process, and export the 4D XCAT images. A robust workflow has been established to implement the 4D XCAT phantom for 4D RT application.


Influence of Atomic Hydrogen, Band Bending, and Defects in the Top Few Nanometers of Hydrothermally Prepared Zinc Oxide Nanorods.
We report on the surface, sub-surface (top few nanometers) and bulk properties of hydrothermally grown zinc oxide (ZnO) nanorods (NRs) prior to and after hydrogen treatment. Upon treating with atomic hydrogen (H*), upward and downward band bending is observed depending on the availability of molecular H2O within the structure of the NRs. In the absence of H2O, the H* treatment demonstrated a cleaning effect of the nanorods, leading to a 0.51 eV upward band bending. In addition, enhancement in the intensity of room temperature photoluminescence (PL) signals due to the creation of new surface defects could be observed. The defects enhanced the visible light activity of the ZnO NRs which were subsequently used to photocatalytically degrade aqueous phenol under simulated sunlight. On the contrary, in the presence of H2O, H* treatment created an electronic accumulation layer inducing downward band bending of 0.45 eV (~1/7th of the bulk ZnO band gap) along with the weakening of the defect signals as observed from room temperature photoluminescence spectra. The results suggest a plausible way of tailoring the band bending and defects of the ZnO NRs through control of H2O/H* species.


Methylnaltrexone bromide for the treatment of opioid-induced constipation in patients with advanced illness--a cost-effectiveness analysis.
Opioid-induced constipation is a common adverse event in patients with advanced illness and has a significant negative impact on patients' quality of life and costs. To examine the cost-effectiveness of treating opioid-induced constipation with methylnaltrexone bromide (MNTX) plus standard care compared with standard care alone in patients with advanced illness who receive long-term opioid therapy from a third-party payer perspective in the Netherlands. A decision-analytical model was created in which advanced-illness patients with constipation were treated with MNTX plus standard care or standard care alone. Clinical efficacy in terms of percentage of patients with rescue-free laxation and time to rescue-free laxation were obtained from a randomized, controlled clinical study. Resource use, costs, utilities and mortality were obtained from published literature and supplemented with data from clinical experts. Treatment with MNTX plus standard care results in more days without constipation symptoms. Cost of MNTX was mostly offset by reduction in other constipation-related costs. Thus, treating with MNTX plus standard care is cost-effective, with an incremental cost per QALY of 40,865 euro. Results were robust to changes in all parameters. Although using MNTX may increase total costs, MNTX plus standard care is cost-effective in treating advanced-illness patients with opioid-induced constipation.


Human Mig chemokine: biochemical and functional characterization.
Mig is a chemokine of the CXC subfamily that was discovered by differential screening of a cDNA library prepared from lymphokine-activated macrophages. The mig gene is inducible in macrophages and in other cells in response to interferon (IFN)-gamma. We have transfected Chinese hamster ovary (CHO) cells with cDNA encoding human Mig and we have derived CHO cell lines from which we have purified recombinant human Mig (rHuMig). rHuMig induced the transient elevation of [Ca2+]i in human tumor-infiltrating T lymphocytes (TIL) and in cultured, activated human peripheral blood-derived lymphocytes. No responses were seen in human neutrophils, monocytes, or Epstein-Barr virus-transformed B lymphoblastoid cell lines. rHuMig was chemotactic for TIL by a modified Boyden chamber assay but rHuMig was not chemotactic for neutrophils or monocytes. The CHO cell lines, IFN-gamma-treated human peripheral-blood monocytes, and IFN-gamma-treated cells of the human monocytic cell line THP-1 all secreted multiple and identical HuMig species as revealed by SDS-PAGE. Using the CHO-derived rHuMig, we have shown that the species' heterogeneity is due to proteolytic cleavage at basic carboxy-terminal residues, and that the proteolysis occurs before and not after rHuMig secretion by the CHO cells. The major species of secreted rHuMig ranged from 78 to 103 amino acids in length, the latter corresponding to the full-length secreted protein predicted from the HuMig cDNA. Carboxy-terminal-truncated forms of rHuMig were of lower specific activity compared to full-length rHuMig in the calcium flux assay, and the truncated species did not block the activity of the full-length species. It is likely that HuMig plays a role in T cell trafficking and perhaps in other aspects of the physiology of activated T cells.


Pharmacovigilance of oral bisphosphonates: adverse effects manifesting in the soft tissue of the oral cavity.
It is well known that oral bisphosphonates can induce necrosis of the osseous structures of the jaws. However, there seems to be a limited awareness that oral bisphosphonates can also induce adverse effects in the soft tissues of the oral cavity, as indicated by the paucity of reported cases in the literature. Because oral bisphosphonates are widely used drugs for several skeletal conditions, it is reasonable to assume that mucosal adverse effects are more common than the small number of published cases indicates. The purpose of this study was to investigate whether this adverse effect of bisphosphonates is represented as reports from health practitioners in an adverse drug reaction database, as well as to gain knowledge about which substances are being associated with adverse drug reactions affecting the oral mucosa. The database of the Medical Products Agency-Sweden was searched for adverse effects from oral bisphosphonates manifesting in the oral and maxillofacial region. Reports of reactions limited to the soft tissues of the oral cavity were selected and further analyzed. Only those reports showing recovery or improvement after the cessation of bisphosphonate use were included in the study. A total of 83 cases of adverse reactions to oral bisphosphonates were retrieved from the search. Of these, 12 were included in the study. They were associated with the use of alendronate, etidronate and risedronate, in descending order. Sixteen percent of the reports comprising the oral and maxillofacial region were limited to the oral mucosa and reported recovery or improvement after discontinuation of the drug. Adverse effects of oral bisphosphonates with manifestations in the soft tissue of the oral cavity seem to be more common than the small number of published cases indicates. However, considering that oral bisphosphonates are widely used drugs, the incidence is still low. These adverse drug reactions are not limited to alendronate and may also be induced by etidronate and risedronate. Still, a significant proportion of the cases are associated with alendronate. Regardless of the substance used, discontinuing the drug is an effective treatment for the mucosal lesions.


The Oregon brain aging study: neuropathology accompanying healthy aging in the oldest old.
To describe the relationship between neuropathologic aging and longitudinal measures of cognitive function in healthy oldest old individuals. Nondemented individuals without cardiovascular or other age-associated diseases of age > or =85 years were followed until death. Regional postmortem measures of senile plaque (SP) and neurofibrillary tangle (NFT) severity were examined in relationship to clinical status, cognitive measures, and rate of cognitive change. Among 19 healthy individuals, 10 became demented or had incipient dementia develop. Clinical status and rate of change in cognitive scores correlated with increasing brain lesion burden, particularly in neocortical regions. Compared to demented individuals, nondemented individuals had few or no neocortical NFT (p = 0.009) or SP (p = 0.001). There was a strong correlation between rate of cognitive change on Mini-Mental State Examination (MMSE) and neocortical NFT (r = 0.859, p = 0.001). The few NFT and SP in nondemented patients had a predilection for limbic areas. These results support a continuum in which AD is infrequent in the healthy, cognitively stable, oldest old. The minimal abnormalities in cognitively stable individuals are consistent with the notion that preclinical pathologic AD precedes obvious cognitive impairment. Longitudinal cognitive testing shows an increased burden of neuropathologic changes in those who have cognitive decline but are not functionally impaired and do not meet criteria for the diagnosis of dementia. The strong relationship between cumulative pathologic changes and rate of cognitive decline suggests that these lesions may have clinical consequences at any age and are not likely to be benign senescent changes.


Genistein and Myd88 Activate Autophagy in High Glucose-Induced Renal Podocytes In Vitro.
BACKGROUND Renal podocyte damage plays a crucial role in the development of diabetic nephropathy. Genistein is derived from a leguminous plant, and MyD88 and TRIF are adaptor molecules in the Toll-like receptor (TLR) signaling pathway, which may play a role in autophagy. In this study, we utilized an in vitro high glucose (HG)-treated podocyte model to investigate the effects and underlying mechanisms of Genistein and MyD88 or TRIF siRNA induced autophagy and renal protection. MATERIAL AND METHODS An immortalized mouse podocyte cell line was treated with HG, Genistein, chloroquine, and/or transfected with specific Myd88 and TRIF siRNAs. The formation of autophagosomes and related autophagic vacuoles were monitored by transmission electron microscopy. The expression of autophagy-related factors and podocyte structure and functional markers, including LC3, p62, p-mTOR, synaptopodin, and nephrin, were measured by Western blot, and LC3 and p-mTOR expression were also assessed by immunofluorescence. RESULTS We showed that HG transiently (after 6-h exposure) induced expression of the autophagy activation marker LC3-II in podocytes. Genistein treatment induced autophagy in both normal and HG-treated podocytes through inactivating mTOR signaling. Moreover, Genistein protected podocytes against chloroquine in HG-cultured conditions in vitro by maintaining the level of autophagy-related proteins. In addition, MyD88 siRNA downregulated expression of autophagy-related proteins, whereas Genistein treatment reversed these effects. CONCLUSIONS This study demonstrated that Genistein-induced autophagy could be a potential treatment strategy for glomerular diseases.


Effects of antiglaucoma drugs on [32P]orthophosphate incorporation into phospholipids of cat iris and ciliary process.
The effects of antiglaucoma drugs on [32P]-orthophosphate incorporation into phospholipids of iris and ciliary process were investigated. Both iris and ciliary process rapidly incorporated 32Pi into the major phospholipids, with the acidic phosphoinositides demonstrating a greater labelling than phosphatidylcholine, indicating a greater turnover. The muscarinic agonists, carbachol and pilocarpine, stimulated 32Pi-labelling of phosphatidylinositol (PI) and phosphatidic acid (PA) in both iris and ciliary process. These effects were blocked by atropine, suggesting that the response was mediated through muscarinic receptors. The beta blocking ocular hypotensive drugs, propranolol, timolol and atenolol, produced varying effects on 32P incorporation into phospholipids of iris and ciliary process. Propranolol stimulated 32Pi-labelling into phosphatidylinositol 4', 5' bisphosphate (PIP2), phosphatidylinositol 4' phosphate (PIP), PI and PA. Timolol decreased 32Pi-incorporation into PIP2 and PI, whereas atenolol, a selective beta 1 antagonist, had no significant effect on 32Pi-labelling of phospholipids. The above findings on propranolol agree with previous observations which demonstrated that propranolol redirects glycerolipid metabolism through multiple effects on the enzymes in phospholipid biosynthesis, particularly in stimulating phosphatidylinositol kinases. The results with timolol suggest that this drug may decrease phosphoinositide hydrolysis. The effects of these ocular hypotensive, non-selective beta blocking drugs on phospholipid turnover may ultimately limit the accumulation of breakdown products which could serve as cellular messengers.


Guidelines of care for vascular lasers and intense pulse light sources from the European Society for Laser Dermatology.
Lasers and non-coherent intense pulse light sources (IPLS) are based on the principle of selective photothermolysis and can be used for the treatment of many vascular skin lesions. A variety of lasers has been developed for the treatment of congenital and acquired vascular lesions which incorporate these concepts into their design. Although laser and light sources are very popular due to their non-invasive nature, caution should be considered by practitioners and patients to avoid permanent side-effects. The aim of these guidelines is to give evidence-based recommendations for the use of lasers and IPLS in the treatment of vascular lesions. These guidelines were produced by a Consensus Panel made up of experts in the field of vascular laser surgery under the auspices of the European Society of Laser Dermatology. Recommendations on the use of vascular lasers and IPLS were made based on the quality of evidence for efficacy, safety, tolerability, cosmetic outcome, patient satisfaction/preference and, where appropriate, on the experts' opinion. The recommendations of these guidelines are graded according to the American College of Chest Physicians Task Force recommendations on Grading Strength of Recommendations and Quality of Evidence in Clinical Guidelines. Lasers and IPLS are very useful and sometimes the only available method to treat various vascular lesions. It is of a paramount importance that the type of laser or IPLS and their specific parameters are adapted to the indication but also that the treating physician is familiar with the device to be used. The crucial issue in treating vascular lesions is to recognize the immediate end-point after laser treatment. This is the single most important factor to ensure both the efficacy of the treatment and avoidance of serious side-effects.


Morphogenesis in robot swarms.
Morphogenesis allows millions of cells to self-organize into intricate structures with a wide variety of functional shapes during embryonic development. This process emerges from local interactions of cells under the control of gene circuits that are identical in every cell, robust to intrinsic noise, and adaptable to changing environments. Constructing human technology with these properties presents an important opportunity in swarm robotic applications ranging from construction to exploration. Morphogenesis in nature may use two different approaches: hierarchical, top-down control or spontaneously self-organizing dynamics such as reaction-diffusion Turing patterns. Here, we provide a demonstration of purely self-organizing behaviors to create emergent morphologies in large swarms of real robots. The robots achieve this collective organization without any self-localization and instead rely entirely on local interactions with neighbors. Results show swarms of 300 robots that self-construct organic and adaptable shapes that are robust to damage. This is a step toward the emergence of functional shape formation in robot swarms following principles of self-organized morphogenetic engineering.


Indomethacin in the treatment of lithium-induced nephrogenic diabetes insipidus.
Nephrogenic diabetes insipidus (NDI) is a frequent complication in patients receiving long-term lithium therapy. Both thiazide diuretics and amiloride may reduce the polyuria, but the use of each is associated with problems. We report the results of a clinical trial using the nonsteroidal anti-inflammatory drug indomethacin to treat a patient with well-documented lithium-induced NDI that persisted following cessation of lithium treatment. The administration of a single dose of indomethacin resulted in a dramatic decrease in urine volume and increase in urine osmolality that persisted for several hours, and was independent of renal hemodynamic changes. Subsequently, the patient experienced a sustained, favorable effect on her polyuria during long-term (3 months) indomethacin therapy without a deleterious effect on her renal function. Indomethacin may be a useful therapeutic tool for the amelioration of lithium-induced NDI.


Current strategies and applications of tissue engineering in dentistry--a review part 1.
A new direction in the field of vital pulp therapy is given by the introduction of tissue engineering as an emerging science. It aims to regenerate a functional tooth-tissue structure by the interplay of three basic key elements: stem cells, morphogens and scaffolds. It is a multidisciplinary approach that combines the principles of biology, medicine, and engineering to repair and/or regenerate a damaged tissue and/or organ. This two part article reviews and discusses the basic concept and strategies so far studied and researched for the engineering of basic dental tissues, to restore a functional tooth anatomy. This first part focuses on a detailed description of key elements used in tissue engineering and their applied clinical applications in dentistry. The second part will deal with the strategies that are being used and/or developed to regenerate the tooth tissues with the help of this scientific principle. The field of tissue engineering has recently shown promising results and a good prospect in dentistry for the development of the most ideal restorations to replace the lost tooth structure with a functional replacement.


Closed-loop phase stepping in a calibrated fiber-optic fringe projector for shape measurement.
Active homodyne feedback control can be used to stabilize an interferometer against unwanted phase drifts introduced by, for example, temperature gradients. The technique is commonly used in fiber-optic sensors to maintain the fiber at its most sensitive (quadrature) position. We describe an extension of the technique to introduce stabilized, pi/2-rad phase steps in a full-field interferometer. The technique was implemented in a single-mode, fiber-optic interference fringe projector used for shape measurement and can be easily applied to other fiber- or bulk-optic interferometers, for example, speckle pattern and holographic interferometers. Fresnel reflections from the distal fiber ends undergo a double pass in the fibers and interfere at the fourth port of a directional coupler. The interference intensity (and hence phase) is maintained at quadrature by feedback control to a phase modulator in one of the fiber arms. Stepping between quadrature positions (separated by pi rad for light undergoing a double pass) introduces stabilized phase steps in the projected fringes (separated by pi/2 rad for a single pass). A root-mean-square phase stability of 0.61 mrad in a 50-Hz bandwidth and phase step accuracy of 1.17 mrad were measured.


A threonine-based targeting signal in the human CD1d cytoplasmic tail controls its functional expression.
CD1d molecules are MHC class I-like molecules that present lipids to a unique subpopulation of T cells called NKT cells. The cytoplasmic tail of human CD1d possesses a tyrosine-based endosomal targeting motif (YXXZ). As such, these molecules traffic through the endocytic pathway, where it is believed that they are loaded with the antigenic lipid that stimulates NKT cells. In the current study, it was found that the T322 residue in the human CD1d tail is a major signal controlling transport to the cell surface and thus its functional expression. Mimicking the phosphorylation of this residue or removal of the entire cytoplasmic tail negates its ability to regulate CD1d trafficking, resulting in lysosomal targeting and degradation. These results demonstrate an important role of a heretofore unknown signal in the cytoplasmic tail of CD1d that may have relevance to other type I integral membrane proteins that traverse through the endocytic pathway.


Ultrastructure of Mycobacterium marinum granuloma in striped bass Morone saxatilis.
An emerging epizootic of mycobacteriosis currently threatens striped bass Morone saxatilis populations in Chesapeake Bay, USA. Several species of mycobacteria, including Mycobacterium marinum, species resembling M. avium, M. gordonae, M. peregrinum, M. scrofulaceum and M. terrae, and the new species M. shottsii have been isolated from diseased and healthy bass. In this study, we describe the ultrastructure of developing M. marinum granulomas in experimentally infected bass over a period of 45 wk. The primary host response to injected mycobacteria was formation of large macrophage aggregations containing phagocytosed bacilli. M. marinum were always contained within phagosomes. Close association of lysosomes with mycobacterial phagosomes, as well as the presence of electron-opaque material within phagosomes, suggested phagolysosomal fusion. Development of granulomas involved epithelioid transformation of macrophages, followed by appearance of central necrosis. Desmosomes were present between mature epithelioid cells. The necrotic core region of M. marinum granulomas was separated from overlying epithelioid cells by several layers of flattened, electron-opaque spindle-shaped cells. These cells appeared to be formed by compression of epithelioid cells and, aside from a flattened nucleus, did not possess recognizable organelles. Following the development of well-defined, paucibacillary granulomas, secondary disease was observed. Recrudescence was marked by bacterial replication followed by disruption of granuloma architecture, including loss of epithelioid and spindle cell layers. In advanced recrudescent lesions, normal tissue was replaced by macrophages, fibroblasts, and other inflammatory leukocytes. Large numbers of mycobacteria were observed, both intracellular and suspended in cellular debris.


Long-Term Use of Everolimus in Patients with Tuberous Sclerosis Complex: Final Results from the EXIST-1 Study.
Everolimus, a mammalian target of rapamycin (mTOR) inhibitor, has demonstrated efficacy in treating subependymal giant cell astrocytomas (SEGAs) and other manifestations of tuberous sclerosis complex (TSC). However, long-term use of mTOR inhibitors might be necessary. This analysis explored long-term efficacy and safety of everolimus from the conclusion of the EXIST-1 study (NCT00789828). EXIST-1 was an international, prospective, double-blind, placebo-controlled phase 3 trial examining everolimus in patients with new or growing TSC-related SEGA. After a double-blind core phase, all remaining patients could receive everolimus in a long-term, open-label extension. Everolimus was initiated at a dose (4.5 mg/m2/day) titrated to a target blood trough of 5-15 ng/mL. SEGA response rate (primary end point) was defined as the proportion of patients achieving confirmed ≥50% reduction in the sum volume of target SEGA lesions from baseline in the absence of worsening nontarget SEGA lesions, new target SEGA lesions, and new or worsening hydrocephalus. Of 111 patients (median age, 9.5 years) who received ≥1 dose of everolimus (median duration, 47.1 months), 57.7% (95% confidence interval [CI], 47.9-67.0) achieved SEGA response. Of 41 patients with target renal angiomyolipomas at baseline, 30 (73.2%) achieved renal angiomyolipoma response. In 105 patients with ≥1 skin lesion at baseline, skin lesion response rate was 58.1%. Incidence of adverse events (AEs) was comparable with that of previous reports, and occurrence of emergent AEs generally decreased over time. The most common AEs (≥30% incidence) suspected to be treatment-related were stomatitis (43.2%) and mouth ulceration (32.4%). Everolimus use led to sustained reduction in tumor volume, and new responses were observed for SEGA and renal angiomyolipoma from the blinded core phase of the study. These findings support the hypothesis that everolimus can safely reverse multisystem manifestations of TSC in a significant proportion of patients. ClinicalTrials.gov NCT00789828.


Renal disease and occupational exposure to organic solvents: a case referent approach.
Several recent studies have suggested that a relation may exist between exposure to occupational organic solvents and diseases of the kidney--particularly malignancy and glomerulonephritis. Two case referent studies were undertaken in the West Midlands to investigate these possibilities. In the case of renal cancer 54 live cases of biopsy proved adenocarcinoma of the kidney were compared with an equal number of community based healthy referents matched for age, sex, place of residence, and socioeconomic and ethnic grouping. For glomerulonephritis, 50 biopsy proved cases were matched in the same manner with 50 referents. Fourteen other patients were also reviewed who, on biopsy, proved not to have glomerulonephritis. For both sets of cases and their referents each individual was interviewed and a detailed account obtained of medical history and environmental exposures. Exposure to solvents was assessed independently and "blind" in a semiquantitative way by an experienced occupational hygienist. Past exposure was estimated for 10 different solvent types and 17 material types. No relation was found between exposure to solvents and renal cancer or glomerulonephritis. In the case of renal cancer the numbers studied only precluded a fourfold excess risk. For glomerulonephritis, the study, although methodologically superior to most other published studies and of similar size, was of similar power to the renal cancer investigation.


Proceedings of the international consensus conference on breast cancer risk, genetics, & risk management, April, 2007.
A consensus conference including thirty experts was held in April, 2007, to discuss risk factors for breast cancer and their management. Four categories of risk were outlined, from breast cancer "average" through "very high" risk, the latter including individuals with high penetrance BRCA1/2 gene mutations. Guidelines for management of patients in each of these categories were discussed, with the major portion of the conference being devoted to individuals with BRCA1/2 mutations. Prevalence of these mutations in the general populations was estimated to be 1 in 250-500 individuals, with an increased prevalence in Ashkenazic Jews and other founder groups. Risk reduction strategies for these individuals include surveillance, with or without chemoprevention drugs, or surgical procedures to remove the organs at risk, i.e., bilateral mastectomy and/or bilateral salpingo-oophorectomy. These risk reduction strategies were evaluated fully, and recommendations were made for the care of patients in each of the risk categories. These guidelines for patient care were approved by the entire group of experts.


[Dual X-ray absorptiometry [DEXA] in normal Chilean women. Correlation between reference values and anthropometric variables].
Bone mineral density results are expressed as the number of standard deviations from reference values in similar populations. However these reference values have not been obtained locally and there may be discrepancies between Chilean and other Hispanic populations. To evaluate the results of DEXA in healthy Chilean women and to correlate them with reference values and anthropometric parameters. We studied prospectively 166 normal women aged between 15 and 55 years, using a Hologic QDR 1000 equipment, yielding bone mineral density (BMD), bone mineral content (BMC) and bone area (A) in the lumbar spine (LS) and femoral neck (FN). These bone densitometric variables vere correlated with weight, height and age of women and compared against the reference values of the Hologic equipment. Relation between BMC and A was evaluated using a regression analysis. BMD and BMC were lower to reference values in most age groups, especially for LS. There was a significant correlation of BMD with A in LS (r: 0.278, p < 0.001), weght in both LS (r: 0.382, p < 0.0001) and FN (r: 0.266, p < 0.001), height in LS (r: 0.258, p < 0.001) and age in FN (r: -0.231, p < 0.01). Only in LS, regression analysis between A and BMC did not show direct proportionality. BMD of healthy Chilean women was lower than the reference values of the Hologic equipment for most age groups. Bone densitometric variables are non uniformly influenced by weight, height and size of bone structures (A) in normal women, especially in LS. Correct interpretation of DEXA requires the validation of reference values and to assess the value of bone area measurements.


Mechanical disintegration of sewage sludge.
Mechanical disintegration can be used for an accelerated and improved anaerobic digestion of excess sludge. The hydrolysis is the limiting step of this process. Mechanical disintegration can be used to disrupt the cell walls and to cause the release of the organic material from the cells. Particle size analysis describes the size reduction but is not suitable for characterising the release of the organic material and the cell disruption. Two biochemical methods were developed for these phenomena. One of the parameters provides information about the disruption of micro-organisms, the other one gives information about the release of organic material. Different ultrasonic homogenizers, a high pressure homogenizer and stirred ball mills were used for disintegration experiments using various parameters. The influences of a mechanical disintegration on the particle size and of the energy intensity on the disintegration were investigated. Further investigations had to detect the influence of the solid content on the disintegration results. For sludge with a higher solid content better results in terms of energy consumption could be achieved. An optimum of the bead diameter and the stress intensity in stirred ball mills could be detected. A comparison of the results of different methods of sludge disintegration shows that the investigated ultrasonic homogenizers are inferior to a high pressure homogenizer and a stirred ball mill in terms of energy consumption.


Jugular bulb temperature: comparison with brain surface and core temperatures in neurosurgical patients during mild hypothermia.
Blood temperature at the jugular bulb was monitored in 10 patients undergoing neurovascular procedures that used induced mild hypothermia, and its correlation with surface brain, core, and peripheral temperatures was determined. The study was motivated by the difficulty encountered in directly measuring global brain temperature and the poor correlations between various core and peripheral sites temperatures and brain temperature, particularly during deep hypothermia. Although not statistically significant, previous studies have suggested a trend toward higher brain temperatures. Temperatures from the jugular bulb (collected using a No. 5 French Swan-Ganz catheter) as well as from subdural, pulmonary artery, esophagus, tympanic membrane, and bladder sites were analyzed during three surgical conditions: prior to incision, with the dura open, and after closure of the dura. No complications related to placement of the jugular bulb catheter, induced hypothermia, or temperature monitoring were seen. The authors found that jugular bulb temperature was similar to pulmonary artery and esophageal temperatures; although prior to incision it tended to be higher than that found at the pulmonary artery, most commonly by 0.2 degrees C. Surface brain temperature was cooler than all other temperatures (p < 0.05), except that of the tympanic membrane, and was particularly sensitive to environmental variations. Finally, as has been shown by others, bladder temperature lagged substantially behind core temperatures particularly during rapid cooling and rewarming of the patient. In summary, monitoring of jugular bulb temperature is a feasible technique, and temperatures measured in the jugular bulb are similar to core temperatures.


Measuring quality of life in stroke subjects receiving an implanted neural prosthesis for drop foot.
The aim was to determine if the treatment of a drop foot by means of an implantable two-channel peroneal nerve stimulator improves health-related quality of life (HRQoL). All subjects were measured at baseline and after a follow-up period of 12 and 26 weeks. Twenty-nine stroke survivors with chronic hemiplegia with drop foot who fulfilled the predefined inclusion and exclusion criteria were included in the present randomized controlled trial. The intervention group received an implantable two-channel peroneal nerve stimulator for correction of their drop foot. The control group continued using their conventional walking device, consisting of an ankle-foot orthosis, orthopedic shoes, or no device. HRQoL was assessed in two different ways: (i) by taking descriptive measures, that is, the Short Form-36 (SF-36; generic measure) and the Disability Impact Profile (DIP; specific measure); and (ii) by obtaining preference-based utilities both measured with the time trade-off (direct way) and by calculating them from the EuroQol (EQ-5D) and SF-36. A significant positive effect of the implantable device was found on the physical functioning domain, the general health domain, and the physical component summary score of the SF-36. For the DIP, a significant improvement was found on the domains mobility, self-care, and psychological status in the intervention group. Regarding the preference-based utility measures, a significant effect was found for the utility index calculated from the EQ-5D. The implantable two-channel peroneal nerve stimulator seems to be efficient to improve HRQoL, mainly the domains related to physical functioning. A relation was present between the utility indexes calculated from the EQ-5D and SF-36.


Protecting human health and security in digital Europe: how to deal with the "privacy paradox"?
This article is the result of an international research between law and ethics scholars from Universities in France and Switzerland, who have been closely collaborating with technical experts on the design and use of information and communication technologies in the fields of human health and security. The interdisciplinary approach is a unique feature and guarantees important new insights in the social, ethical and legal implications of these technologies for the individual and society as a whole. Its aim is to shed light on the tension between secrecy and transparency in the digital era. A special focus is put from the perspectives of psychology, medical ethics and European law on the contradiction between individuals' motivations for consented processing of personal data and their fears about unknown disclosure, transferal and sharing of personal data via information and communication technologies (named the "privacy paradox"). Potential benefits and harms for the individual and society resulting from the use of computers, mobile phones, the Internet and social media are being discussed. Furthermore, the authors point out the ethical and legal limitations inherent to the processing of personal data in a democratic society governed by the rule of law. Finally, they seek to demonstrate that the impact of information and communication technology use on the individuals' well-being, the latter being closely correlated with a high level of fundamental rights protection in Europe, is a promising feature of the socalled "e-democracy" as a new way to collectively attribute meaning to large-scale online actions, motivations and ideas.


Extent of apoptosis in ovarian serous carcinoma: relation to mitotic and proliferative indices, p53 expression, and survival.
To assess the extent of apoptosis in ovarian serous carcinoma and to examine possible relations between apoptosis, cell proliferation, p53 overexpression, and patient survival. Apoptotic and mitotic indices were obtained by examining haematoxylin and eosin stained sections from 30 patients with ovarian serous carcinoma. Apoptosis was also evaluated semiquantitatively by in situ end labelling of fragmented DNA. Expression of p53 and determination of Ki-67 labelling indices were based on immunohistochemical staining. Clinical details were obtained from patients' clinical records. For statistical analysis, Fisher's exact test, parametric (Pearson) linear correlations, and the Kaplan-Meier method were used. The mean apoptotic index was 1.3% (range 0.02-3.9%), the mean mitotic index was 0.4% (range 0.02-1.1%), and the mean Ki-67 labelling index was 16% (range 4-32%). There were significant correlations between the apoptotic and mitotic indices (p < 0.0205) and between the mitotic and Ki-67 labelling indices (p < 0.024). There was a significant correlation between a high apoptotic index and poor prognosis (p < 0.02). p53 was overexpressed in 16 cases but the extent of apoptosis and outcome were both independent of p53 status. These results suggest that regulation of apoptosis is an integral component of tumour cell kinetics in ovarian serous carcinoma, and that increased apoptosis is indicative of aggressive tumour growth. p53 expression did not correlate with altered apoptosis, but the possibility of an attenuated apoptotic response to subsequent DNA damage by anticancer agents is not excluded.


C-reactive protein-guided antibiotic prescribing for COPD exacerbations: a qualitative evaluation.
Antibiotics are prescribed to >70% of patients presenting in primary care with an acute exacerbation of chronic obstructive pulmonary disease (AECOPD). The PACE randomised controlled trial found that a C-reactive protein point-of-care test (CRP-POCT) management strategy for AECOPD in primary care resulted in a 20% reduction in patient-reported antibiotic consumption over 4 weeks. To understand perceptions of the value of CRP-POCT for guiding antibiotic prescribing for AECOPD; explore possible mechanisms, mediators, and pathways to effects; and identify potential barriers and facilitators to implementation from the perspectives of patients and clinicians. Qualitative process evaluation in UK general practices. Semi-structured telephone interviews with 20 patients presenting with an AECOPD and 20 primary care staff, purposively sampled from the PACE study. Interviews were audio-recorded, transcribed, and analysed using framework analysis. Patients and clinicians felt that CRP-POCT was useful in guiding clinicians' antibiotic prescribing decisions for AECOPD, and were positive about introduction of the test in routine care. The CRP-POCT enhanced clinician confidence in antibiotic prescribing decisions, reduced decisional ambiguity, and facilitated communication with patients. Some clinicians thought the CRP-POCT should be routinely used in consultations for AECOPD; others favoured use only when there was decisional uncertainty. CRP-POCT cartridge preparation time and cost were potential barriers to implementation. CRP-POCT-guided antibiotic prescribing for AECOPD had high acceptability, but commissioning arrangements and further simplification of the CRP-POCT need attention to facilitate implementation in routine practice.


A daily diary study on adolescent emotional experiences: Measurement invariance and developmental trajectories.
Adolescence is an important time for emotional development. Recently, daily diary methods are increasingly employed in research on emotional development and are used to explore the development of and sex differences in emotions during adolescence. However, before drawing conclusions about sex differences and developmental trends, one needs to ensure that the same construct is measured across sex and time. The present study tested measurement invariance of daily emotion assessments across sex, short-term (days within weeks) and long-term periods (days across years) in a sample of 394 adolescents (55.6% male) that were followed from ages 13 to 18. Moreover, the study examined the developmental trajectories of adolescent emotional experiences. Adolescents rated their daily emotions (happiness, anger, sadness, anxiety) during each day of a normal school week (Monday to Friday) for 3 weeks per year for 5 years (i.e., 15 weeks × 5 days = 75 assessments in total). Measurement invariance analyses suggest that the measurement of adolescent daily mood was invariant between boys and girls and across shorter and longer time intervals. Moreover, latent growth curve analyses showed that happiness decreased from early to middle adolescence, whereas anger, sadness, and anxiety increased. Anger returned to baseline toward late adolescence. In contrast, the decrease of happiness and the increase of anxiety leveled off without reversing, whereas sadness continued to increase. The discussion highlights the implications of measurement invariance in research on individual and developmental differences and discusses the findings in light of normative emotional development. (PsycINFO Database Record


Translational control of phloem development by RNA G-quadruplex-JULGI determines plant sink strength.
The emergence of a plant vascular system was a prerequisite for the colonization of land; however, it is unclear how the photosynthate transporting system was established during plant evolution. Here, we identify a novel translational regulatory module for phloem development involving the zinc-finger protein JULGI (JUL) and its targets, the 5' untranslated regions (UTRs) of the SUPPRESSOR OF MAX2 1-LIKE4/5 (SMXL4/5) mRNAs, which is exclusively conserved in vascular plants. JUL directly binds and induces an RNA G-quadruplex in the 5' UTR of SMXL4/5, which are key promoters of phloem differentiation. We show that RNA G-quadruplex formation suppresses SMXL4/5 translation and restricts phloem differentiation. In turn, JUL deficiency promotes phloem formation and strikingly increases sink strength per seed. We propose that the translational regulation by the JUL/5' UTR G-quadruplex module is a major determinant of phloem establishment, thereby determining carbon allocation to sink tissues, and that this mechanism was a key invention during the emergence of vascular plants.


Optimal Fourier control performance and speckle behavior in high-contrast imaging with adaptive optics.
High-contrast imaging with adaptive optics (AO) for planet detection requires a sophisticated AO control system to provide the best possible performance. We evaluate the performance improvements in terms of residual error and point-spread function intensity provided by optimal Fourier control using detailed end-to-end simulation. Intensity, however, is not the final measure of system performance. We explore image contrast through analysis and simulation results, showing that speckles caused by atmospheric errors behave very differently in a temporal fashion from speckles caused by wavefront sensor noise errors.


In vivo effects of angiotensin II on vascular smooth muscle contraction and blood pressure are mediated through a protein tyrosine-kinase-dependent mechanism.
The effects of in vivo angiotensin II (AII) treatment on protein tyrosine kinase (PTK) levels were examined. It was found that administration of AII to normotensive Wistar-Kyoto rats induced tyrosine phosphorylation in aorta in a time-dependent manner. There was a rapid increase in phosphotyrosine content as early as 1 min, with peak phosphorylation occurring between 5 and 10 min. The response was also dose dependent, with increases in phosphorylation levels from 1 to 1000 micrograms/kg AII. Tyrosine phosphorylation was blocked using the angiotensin type 1 receptor antagonist losartan (10 mg/kg), suggesting that the effects are receptor mediated. Tyrphostin-25 (100 microM), a selective inhibitor of PTKs, when given in vivo was also able to attenuate phosphorylation by AII, further suggesting a PTK-mediated event. To couple these biochemical changes with physiological events, we also examined the ability of AII to induce vasoconstriction and raise systolic blood pressure through a PTK-mediated mechanism. In addition to increasing phosphorylation levels, AII caused a rise in systolic pressure in vivo and induced contraction in vitro. Both of these responses could be attenuated by pretreatment with losartan or tyrphostin-25. This is the first demonstration of the effects of AII on tyrosine phosphorylation in vivo. The data suggest that AII may induce a pressor response at least in part through activation of PTKs and subsequent phosphorylation of smooth muscle contractile proteins or activation of other protein kinases.


'We needed to change the mission statement of the marriage': biographical disruptions, appraisals and revisions among couples living with endometriosis.
The concept of biographical disruption has been widely applied in sociological explorations of chronic illness and has been subject to much theoretical scrutiny, reflection and development. However, little attention has been given to the impact of biographical disruption beyond the individual level. This article explores the concept from a dyadic perspective, utilising data from an exploratory, qualitative study (ENDOPART) that investigated the impact of endometriosis on women and their male partners. In total, 22 couples participated in in-depth, semi-structured, face-to-face interviews. The women and their partners were interviewed separately and, in most cases, simultaneously, by different interviewers. Data analysis was informed by an interpretivist relational approach, foregrounding the meanings participants applied to their experiences, treating interviews as accounts, and exploring partners' accounts in relation to one another. Two analytic approaches generated several themes for exploration in the context of the concept of biographical disruption: sex and intimacy; planning for and having children; working lives and social lives. The article argues that biographical disruptions are social and inter-relational processes and discusses how couples living with endometriosis negotiated these disruptions, how they were appraised and how lives and expectations were revised as a result.


Effect of Bevacizumab vs Aflibercept on Visual Acuity Among Patients With Macular Edema Due to Central Retinal Vein Occlusion: The SCORE2 Randomized Clinical Trial.
Studies have established the efficacy and safety of aflibercept for the treatment of macular edema due to central retinal vein occlusion. Bevacizumab is used off-label to treat this condition despite the absence of supporting data. To investigate whether bevacizumab is noninferior to aflibercept for the treatment of macular edema secondary to central retinal or hemiretinal vein occlusion. The SCORE2 randomized noninferiority clinical trial was conducted at 66 private practice or academic centers in the United States, and included 362 patients with macular edema due to central retinal or hemiretinal vein occlusion who were randomized 1:1 to receive aflibercept or bevacizumab. The first participant was randomized on September 17, 2014, and the last month 6 visit occurred on May 6, 2016. Analyses included data available as of December 30, 2016. Eyes were randomized to receive intravitreal injection of bevacizumab (1.25 mg; n = 182) or aflibercept (2.0 mg; n = 180) every 4 weeks through month 6. The primary outcome was mean change in visual acuity (VA) letter score (VALS) from the randomization visit to the 6-month follow-up visit, based on the best-corrected electronic Early Treatment Diabetic Retinopathy Study VALS (scores range from 0-100; higher scores indicate better VA). The noninferiority margin was 5 letters, and statistical testing for noninferiority was based on a 1-sided 97.5% confidence interval. Among 362 randomized participants (mean [SD] age, 69 [12] years; 157 [43.4%] women; mean [SD] VALS at baseline, 50.3 [15.2] [approximate Snellen VA 20/100]), 348 (96.1%) completed the month 6 follow-up visit. At month 6, the mean VALS was 69.3 (a mean increase from baseline of 18.6) in the bevacizumab group and 69.3 (a mean increase from baseline of 18.9) in the aflibercept group (model-based estimate of between-group difference, -0.14; 97.5% CI, -3.07 to ∞; P = .001 for noninferiority), meeting criteria for noninferiority. Ocular adverse events in the aflibercept group included 4 participants with intraocular pressure (IOP) more than 10 mm Hg greater than baseline; ocular adverse events in the bevacizumab group included 1 participant with endophthalmitis (culture negative), 9 with IOP more than 10 mm Hg greater than baseline, 2 with IOP higher than 35 mm Hg, and 1 with angle-closure glaucoma not attributed to the study drug or procedure. Among patients with macular edema due to central retinal or hemiretinal vein occlusion, intravitreal bevacizumab was noninferior to aflibercept with respect to visual acuity after 6 months of treatment.


A comparative study of experimental optimization and response surface optimization of Cr removal by emulsion ionic liquid membrane.
A comparative study on the optimization of process parameters of an emulsion ionic liquid membrane (EILM) by experimental work and response surface methodology (RSM) has been carried out. EILM was prepared by using kerosene as solvent, Span 80 as surfactant, NaOH as internal reagent, a hydrophobic ionic liquid 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([BMIM](+)[NTf(2)](-)) as a stabilizer and a second ionic liquid tri-n-octylmethylammonium chloride (TOMAC) as a carrier. The prepared EILM was used to separate and concentrate Cr from wastewaters. The comparison between the experimentally optimized and the RSM optimized values was accomplished by optimizing the following parameters: homogenization speed, carrier concentration, internal phase concentration, agitation speed, treat ratio, internal to membrane phase ratio, surfactant concentration and pH of the feed phase. The comparison showed that all the values were in good agreement except for the internal phase concentration and the treat ratio. It was observed that the stability provided by [BMIM](+)[NTf(2)](-) decreased as the extraction progressed due to its high density. Nevertheless, a good stability could be obtained by the combination of [BMIM](+)[NTf(2)](-) and Span 80 during extraction process.


Antenatal consent in the SUPPORT trial: challenges, costs, and representative enrollment.
The purposes of this study were to quantify the time and effort involved in obtaining prenatal consent for the Neonatal Research Network Surfactant Positive Airway Pressure and Pulse Oximetry Randomized Trial (SUPPORT) and to determine whether the enrolled infants were representative of the eligible population. Eligible subjects were likely to deliver in the SUPPORT gestational age window (24-27(6/7)] weeks). Data included who approached the subjects for consent, how often they approached, the duration of each contact, whether consent was obtained, and whether subjects were enrolled in the trial. Eligible, nonenrolled infants entered into the Neonatal Research Network Generic Database throughout the period of SUPPORT enrollment were compared with enrolled infants. A total of 2826 women were identified at 18 sites, 2228 were approached for consent, and 1219 (54.7%) agreed. For 76.9% of those approached, <3 visits (mean: 2.0 +/- 1.2 visits) were required to complete the consent process. Of the 659 infants with consent who were delivered within the study window, 611 were enrolled. Mothers who received a neonatal consultation were more likely to give consent (P < .001). The proportion of infants not exposed to steroids was significantly greater in the nonapproached group than in the approached group (20.0% vs 3.4%; P < .0001). In a trial that involved preterm infants and required prenatal consent, >5 women were identified as being likely to deliver in the SUPPORT gestational age window for each 1 who delivered an enrolled infant.


Air pollution, a possible risk factor for multiple sclerosis.
Studies focusing attention on the effects of environmental pollution on the etiology of multiple sclerosis (MS) are on the increase. The aim of this study was to determine MS prevalence in a city home to an iron and steel factory which causes air pollution. The study was designed as a cross-sectional, population-based, descriptive epidemiologic study. Ereğli city, which has an iron and steel factory and proven air pollution, was screened. Additionally, Devrek city, which is a rural and clean city, located 40 km away from Ereğli was assigned and results were compared. A validated questionnaire was used for screening. McDonald 2010 criteria were used to diagnose cases. 32 261 people were screened in Ereğli, and 21 963 people were screened in Devrek. In total, 41 patients were diagnosed with clinical definite MS. Crude prevalence was found to be 96.1/100 000 in Ereğli and 45.5/100 000 in Devrek. The mean age of patients was 39.8, and the female/male ratio was 1.9. The results of this study indicate a more than double MS prevalence rate in the area home to an iron and steel factory when compared to the rural city. This supports the hypothesis that air pollution may be a possible etiological factor in MS.


Therapy of bacterial sepsis, meningitis and otitis media in infants and children: 1992 poll of directors of programs in pediatric infectious diseases.
To determine current opinions among experts in pediatric infectious diseases for treatment of bacterial sepsis, meningitis and acute otitis media, we polled directors of training programs in January, 1992. Responses were received from 69 centers in the United States and Canada. For initial treatment of presumed bacterial meningitis, the third generation cephalosporins alone or combined with ampicillin have become drugs of choice in all age groups. Most infectious disease programs include dexamethasone in the management of presumed bacterial meningitis for children 2 months of age and older. Third generation cephalosporins are also drugs of choice for presumed sepsis: combined with ampicillin for infants 5 weeks of age; used alone for children 5 months and 12 years of age. Amoxicillin remains the preferred drug for initial treatment of acute otitis media. The combination of amoxicillin and clavulanic acid is favored in the setting of an increased proportion of beta-lactamase-producing bacterial pathogens. Comparison of these results with polls in 1987 and 1989 indicates a shift in recommendations of therapy of presumed bacterial sepsis and meningitis from ampicillin alone or combined with an aminoglycoside or chloramphenicol to use of a third generation cephalosporin alone or combined with ampicillin.


SU-E-T-437: Interfractional Dosimetric Verification of Lung Patients Treated by Passive Double Scattering Proton Radiotherapy.
Proton radiotherapy, with the ability to confine the dose at desired depth, can potentially benefit lung tumor patients by significantly sparing the healthy lung tissue. However, the superior proton dose distribution could be affected by tumor shrinkage due to the quick response and by motion especially related to the respiration. Thus the treatment should be frequently verified and be adjusted accordingly if necessary to achieve the initial treatment goal. A cohort of 20 patients were selected from lung patients treated with passive proton radiotherapy. All those patients were evaluated via 4D-CT scans and found to have tumor motion less than 1 cm. The internal target volumes (ITV) were derived based on the full inspiration and expiration phases. The average of the 4D-CT scan, full inspiration and expiration phases were used for the initial treatment planning. The planning objective was 95% of the prescription dose to at least 95% volume of the ITV. Bi-weekly verification 4D-CT scans were performed to assess the robustness of the initial treatment plan and no replanning was required for target dose variations less then 3%. Compared with the initial treatment plan, the standard deviations of target coverage on inspiration, expiration, and average verification CT scans are within 3% for all the patients, with the maximum difference up to 7%. No statistically significant differences were found among the initial and verification plans (p>0.1). The percentage deviations of OAR sparing were highly variable, e.g., up to 40% for mean lung dose, 100% for mean heart dose, 50% for max cord dose, particularly for OARs receiving small amount of doses. However, the absolute dose deviations are all with OAR's tolerance. Overall, the passive double scattering proton modality allows for robust proton treatment planning and delivery to treat the lung tumors with limited motion.


Tanshinone IIA prevents uric acid nephropathy in rats through NF-κB inhibition.
The purpose of this research is to investigate the effect of tanshinone IIA, an extract of the Chinese medicine Que Xie Hua Yu Tang, on uric acid nephropathy (UAN) and to elucidate the underlying mechanisms. UAN rat model was established. Fifty UAN rats were randomly allocated into 5 groups: adenine-treated group, allopurinol-treated group, and low/middle/high dose of tanshinone IIA-treated groups. Meanwhile, another 10 rats were used as normal controls. Serum uric acid (UA), blood urea nitrogen (BUN), serum creatinine (Scr), MCP-1, and IL-1β levels were measured. Histological staining was performed. Comparison between the adenine group and treatment (allopurinol and tanshinone IIA) groups showed compound treatment could attenuate the inflammation status of the kidneys and decrease serum UA levels. Among different kinds of medicine, tanshinone IIA had similar effects as allopurinol and exerted anti-inflammatory and renal protective effect in a dose-dependent manner. Furthermore, we found tanshinone IIA alone could also inhibit urate-induced MCP-1 and IL-1β overexpression both in vivo and in vitro, accompanied with inhibition of NF-κB translocation from cytosome to nucleus. Tanshinone IIA could protect rats from uric acid-induced kidney damage, probably by attenuating renal inflammatory status.


Valnoctamide as a valproate substitute with low teratogenic potential in mania: a double-blind, controlled, add-on clinical trial.
Valproic acid's well-known teratogenicity limits its use in women of childbearing age. Valnoctamide is an analog of valproate that does not undergo biotransformation to the corresponding free acid. In mice, valnoctamide has been shown to be distinctly less teratogenic than valproate. Valnoctamide is an anticonvulsant, and we hypothesized that valnoctamide is antimanic. We performed a double-blind, five-week, add-on, controlled trial of valnoctamide in mania. Patients were treated with risperidone at doses of the physician's discretion. Valnoctamide or placebo was begun at doses of 600 mg/day and increased to 1200 mg after four days. Weekly ratings by a psychiatrist blind to the study drug were conducted using the Brief Psychiatric Rating Scale (BPRS), the Young Mania Rating Scale (YMRS), and the Clinical Global Impression (CGI). Fifteen valnoctamide patients and 17 placebo patients completed at least one post-baseline week and were included in data analysis. In all efficacy measures valnoctamide was more effective than placebo as an add-on to risperidone, using two-way analysis of variance (ANOVA) with time as the within-subject factor. Two-way ANOVA showed a significant effect of time (p < 0.001) and significant interaction between treatment and time (YMRS: p = 0.012; BPRS: p = 0.007; CGI: p = 0.003). Differences between valnoctamide and placebo were significant from week 3 to week 5. Valnoctamide could be an important valproate substitute for women of childbearing age with bipolar disorder who may become pregnant.


Image-based motion detection: using the concept of weighted directional descriptors.
The use of image guidance in medical applications is constantly growing because of its tremendous impact on the future of health care. Although image-based tissue tracking has been thoroughly explored in the academic literature for years, it has not yet matured to become widely accepted by clinicians. Undetected tissue movements in image-based clinical procedures may cause safety and efficacy difficulties. We introduce an image-based approach for detecting tissue movements during clinical procedures. Our method has been validated in more than 600 true clinical cases. The results show that our algorithm agrees with an expert analysis in 98% of the cases, showing zero events of false alarms and zero events of undetected motion. The results show that the approach provides a clinically ready motion detection algorithm. These robust results are achieved by introducing the concept of weighted directional descriptors (WDDs). The technique analyzes the directivity and confidence level of each anatomical feature and uses it to weight local inputs resulting in a robust motion vector. The robustness is further increased by a novel preprocess that screens out features that may be misleading or are repeated in the adjacent search zone. The technique meets the requirements, as defined by our clinicians, and is now integrated in true medical systems. In particular, our approach has been uniquely developed and integrated into a clinical product. ExAblate is the first Food and Drug Administration (FDA)-approved magnetic resonance (MR)-guided noninvasive surgical device using focused ultrasound therapy. It is used in commercial clinics and in leading medical academic research institutions, attesting to the success of our method and its practical clinical value.


Hibiscus sabdariffa L. tea (tisane) lowers blood pressure in prehypertensive and mildly hypertensive adults.
In vitro studies show Hibiscus sabdariffa L., an ingredient found in many herbal tea blends and other beverages, has antioxidant properties, and, in animal models, extracts of its calyces have demonstrated hypocholesterolemic and antihypertensive properties. Our objective in this study was to examine the antihypertensive effects of H. sabdariffa tisane (hibiscus tea) consumption in humans. A randomized, double-blind, placebo-controlled clinical trial was conducted in 65 pre- and mildly hypertensive adults, age 30-70 y, not taking blood pressure (BP)-lowering medications, with either 3 240-mL servings/d of brewed hibiscus tea or placebo beverage for 6 wk. A standardized method was used to measure BP at baseline and weekly intervals. At 6 wk, hibiscus tea lowered systolic BP (SBP) compared with placebo (-7.2 +/- 11.4 vs. -1.3 +/- 10.0 mm Hg; P = 0.030). Diastolic BP was also lower, although this change did not differ from placebo (-3.1 +/- 7.0 vs. -0.5 +/- 7.5 mm Hg; P = 0.160). The change in mean arterial pressure was of borderline significance compared with placebo (-4.5 +/- 7.7 vs. -0.8 +/- 7.4 mm Hg; P = 0.054). Participants with higher SBP at baseline showed a greater response to hibiscus treatment (r = -0.421 for SBP change; P = 0.010). No effects were observed with regard to age, gender, or dietary supplement use. These results suggest daily consumption of hibiscus tea, in an amount readily incorporated into the diet, lowers BP in pre- and mildly hypertensive adults and may prove an effective component of the dietary changes recommended for people with these conditions.


From immune to olfactory expression: neofunctionalization of formyl peptide receptors.
Variations in gene expression patterns represent a powerful source of evolutionary innovation. In a rodent living about 70 million years ago, a genomic accident led an immune formyl peptide receptor (FPR) gene to hijack a vomeronasal receptor regulatory sequence. This gene shuffling event forced an immune pathogen sensor to transition into an olfactory chemoreceptor, which thus moved from sensing the internal world to probing the outside world. We here discuss the evolution of the FPR gene family, the events that led to their neofunctionalization in the vomeronasal organ and the functions of immune and vomeronasal FPRs.


Characteristics of BDNF-induced weight loss.
There is evidence that central infusion of brain-derived neurotrophic factor (BDNF) induces weight loss in rats. We have begun to investigate the physiological basis for BDNF-induced weight loss by assessing its relationship to (a) appetite, (b) serum indices of metabolic and renal toxicity, and (c) brain monoamine activity in areas associated with feeding or motor function. BDNF (0-6 microgram/day) was infused into the lateral ventricle (LV) of male Long-Evans rats for 14 days. Body weight and food intake were monitored throughout infusion and recovery periods. BDNF induced severe, dose-dependent appetite suppression and weight loss. Although appetite began to recover after the 10th infusion day, body weight had not returned to control values at the end of the recovery period. The weight loss observed in BDNF-infused rats was related to appetite suppression, since uninfused rats that were pair-fed to high dose BDNF-treated rats showed comparable weight loss. Despite severe weight loss, serum BUN, creatinine, thyroxine, glucose, and total protein were not affected by BDNF infusion. Striatal DO-PAC/DA was similarly unaffected by BDNF. In contrast, BDNF-infused rats showed a dose-dependent increase in hypothalamic 5-HIAA/5-HT that was not observed in pair-fed rats, suggesting that the observed increase in hypothalamic 5-HIAA/5-HT was a direct effect of BDNF infusion rather than a secondary effect of food restriction. These data suggest that BDNF may induce appetite suppression and weight loss through a central mechanism.


Focused Directed Evolution of Aryl-Alcohol Oxidase in Saccharomyces cerevisiae by Using Chimeric Signal Peptides.
Aryl-alcohol oxidase (AAO) is an extracellular flavoprotein that supplies ligninolytic peroxidases with H2O2 during natural wood decay. With a broad substrate specificity and highly stereoselective reaction mechanism, AAO is an attractive candidate for studies into organic synthesis and synthetic biology, and yet the lack of suitable heterologous expression systems has precluded its engineering by directed evolution. In this study, the native signal sequence of AAO from Pleurotus eryngii was replaced by those of the mating α-factor and the K1 killer toxin, as well as different chimeras of both prepro-leaders in order to drive secretion in Saccharomyces cerevisiae. The secretion of these AAO constructs increased in the following order: preproα-AAO > preαproK-AAO > preKproα-AAO > preproK-AAO. The chimeric preαproK-AAO was subjected to focused-directed evolution with the aid of a dual screening assay based on the Fenton reaction. Random mutagenesis and DNA recombination was concentrated on two protein segments (Met[α1]-Val109 and Phe392-Gln566), and an array of improved variants was identified, among which the FX7 mutant (harboring the H91N mutation) showed a dramatic 96-fold improvement in total activity with secretion levels of 2 mg/liter. Analysis of the N-terminal sequence of the FX7 variant confirmed the correct processing of the preαproK hybrid peptide by the KEX2 protease. FX7 showed higher stability in terms of pH and temperature, whereas the pH activity profiles and the kinetic parameters were maintained. The Asn91 lies in the flavin attachment loop motif, and it is a highly conserved residue in all members of the GMC superfamily, except for P. eryngii and P. pulmonarius AAO. The in vitro involution of the enzyme by restoring the consensus ancestor Asn91 promoted AAO expression and stability.


Ebolavirus VP35 is a multifunctional virulence factor.
Ebola virus (EBOV) is a member of the filoviridae family that causes severe hemorrhagic fever during sporadic outbreaks, and no approved treatments are currently available. The multifunctional EBOV VP35 protein facilitates immune evasion by antagonizing antiviral signaling pathways and is important for viral RNA synthesis. In order to elucidate regulatory mechanisms and to develop countermeasures, we recently solved the structures of the Zaire and Reston EBOV VP35 interferon inhibitory domain (IID) in the free form and of the Zaire EBOV VP35 IID bound to dsRNA. Together with biochemical, cell biological, and virological studies, our structural work revealed that distinct regions within EBOV VP35 IID contribute to virulence through host immune evasion and viral RNA synthesis. Here we summarize our recent structural and functional studies and discuss the potential of multifunctional Ebola VP35 as a therapeutic target.


Hematopoietic bone marrow cells participate in endothelial, but not epithelial or mesenchymal cell renewal in adult rats.
The extent to which bone marrow (BM) contributes to physiological cell renewal is still controversial. Using the marker human placental alkaline phosphatase (ALPP) which can readily be detected in paraffin and plastic sections by histochemistry or immunohistochemistry, and in ultrathin sections by electron microscopy after pre-embedding staining, we examined the role of endogenous BM in physiological cell renewal by analysing tissues from lethally irradiated wild-type inbred Fischer 344 (F344) rats transplanted (BMT) with unfractionated BM from ALPP-transgenic F344 rats ubiquitously expressing the marker. Histochemical, immunohistochemical and immunoelectron microscopic analysis showed that the proportion of ALPP(+) capillary endothelial cells (EC) profoundly increased from 1 until 6 months after BMT in all organs except brain and adrenal medulla. In contrast, pericytes and EC in large blood vessels were ALPP(-) . Epithelial cells in kidney, liver, pancreas, intestine and brain were recipient-derived at all time-points. Similarly, osteoblasts, chondrocytes, striated muscle and smooth muscle cells were exclusively of recipient origin. The lack of mesenchymal BM-derived cells in peripheral tissues prompted us to examine whether BMT resulted in engraftment of mesenchymal precursors. Four weeks after BMT, all haematopoietic BM cells were of donor origin by flow cytometric analysis, whereas isolation of BM mesenchymal stem cells (MSC) failed to show engraftment of donor MSC. In conclusion, our data show that BM is an important source of physiological renewal of EC in adult rats, but raise doubt whether reconstituted irradiated rats are an apt model for BM-derived regeneration of mesenchymal cells in peripheral tissues.


A comparision of the yield of three tuberculosis screening modalities among people living with HIV: a retrospective quasi-experiemental study.
The Intensified Case Finding (ICF) tool was approved for TB screening in 2011; however there is still paucity of robust data comparing yields of the different ICF screening modalities. We compared yields of three different screening modalities for TB among Patients Living with HIV (PLHIV) in Uganda in order to inform National TB Programs on the most effective TB screening method. This was a retrospective quasi-experimental study conducted at an Out-Patient HIV/AIDS clinic in Uganda. We set out to determine yields of three different TB screening modalities at three time periods: 2006/07 where Passive Case Finding (PCF) was used. Here, no screening questions were administered; the clinician depended on the patient's self report. In 2008/09 embedded Intensified Case Finding Tool (e-ICF) was used; here a data capture field was added to the patient clinical encounter forms to compel clinicians to screen for TB symptoms. In 2010/11 Independent Intensified Case Finding Tool (i-ICF) was used; here a screening data collection form, was used, it had the same screening questions as e-ICF. Routine clinical data, including TB status, were collected and entered into an electronic clinical care database. Analysis was done in STATA and the main outcome estimated was the proportional yield of TB cases for each screening modality. The overall yield of TB cases was 11.18 % over the entire period of the study (2006 - 2011). The intervention-specific yields were 1.86 % for PCF, 14.95 % for e-ICF and 12.47 % for i-ICF. Use of either e-ICF (OR: 9.2, 95 % CI: 4.81-17.73) or i- ICF (OR: 7.7, 95 % CI: 4.02-14.78) significantly detected more TB cases compared to PCF (P <0.001). While the yields of the Active Case Finding modalities (e-ICF & i-ICF) were not significantly different (OR: 0.98, 95 % CI 0.76-1.27, P = 0.89). The active screening modalities (e-ICF & i-ICF) had a comparable TB yield and were eight to nine times more efficient in identifying TB cases when compared to the PCF. Cost effectiveness studies would be informative.