Episodic occurrence of antibodies against the bovine leukemia virus Rex protein during the course of infection in sheep.
Infection by bovine leukemia virus (BLV) is characterized by a long clinical latency after which some individuals develop B-cell tumors. The contributions of the viral regulatory proteins Tax and Rex during clinical latency and disease are incompletely understood. To learn about Rex expression in the host, we used a sensitive immunoprecipitation assay to detect Rex antibodies throughout the course of BLV infection in sheep. Sixty percent of the infected animals produced Rex antibodies in intermittent episodes. This pattern differed markedly from that of antibodies to virion structural proteins, which were maintained in all animals throughout infection. Only one of two animals that developed tumors had detectable Rex antibodies at the time, although the other had previously demonstrated an especially strong Rex antibody response. We examined the Rex response in the context of BLV infection by comparing it with the frequency of circulating mononuclear blood cells that could transcribe BLV RNA or produce infectious virus. Episodes of Rex antibody occurrence followed some but not all increases in the number of BLV-transcribing cells. Since the appearance of circulating antibodies requires that the intracellular Rex protein be available to serve as antigen, the episodic pattern of occurrence of Rex antibodies could result from intermittent killing by virus-specific cytotoxic cells. Fluctuations in titer that were observed during some episodes of Rex response could be due to antibody retention by antigen present in lymphoid tissue.


Fluorescence quenching by polystyrene microspheres in UV-visible and NIR tissue-simulating phantoms.
Tissue-simulating phantoms are widely used for controlled studies of photon transport in turbid media. Here, we describe how polystyrene microspheres, which are often used to simulate optical scattering in such phantoms, can reduce fluorophore quantum yield via collisional quenching. We report studies on UV-visible (fluorescein-based) and NIR (IR125-based) phantoms with differing fluorophore and scatterer concentrations, as well as differing microsphere sizes. Results consistent with the Stern-Volmer relation suggest that the fluorophore intrinsic excited-state lifetime decreased due to collisional quenching from polystyrene microspheres and that the quenching efficiency was dependent on the concentration ratio of fluorophores to microspheres. Lifetime decreases ranging from 10-35% (20%) were measured for fluorescein-based (IR 125-based) phantoms. Since polystyrene microspheres are commonly used in tissue-simulating phantoms for quantitative studies of fluorescence light propagation, their quenching effects on fluorescence intensities may be difficult to separate from intensity losses attributed to optical absorption and scattering in the phantom unless fluorescence lifetime measurements are performed simultaneously.


Client Perspectives on Reclaiming Participation After a Traumatic Spinal Cord Injury in South Africa.
The development of rehabilitation services promoting participation in people living with a traumatic spinal cord injury (TSCI) is of major concern for physical therapists. What the client sees as effective participation, barriers, and facilitators might be different due to their particular context. This study was conducted to gain insight into the experiences of attaining an important outcome in a developing context. The aim of this study was to explore the experiences of reclaiming participation in community-dwelling people with TSCI in South Africa. This was an explorative, qualitative study with inductive content analysis. Individual semistructured interviews were conducted with 17 adults living with a TSCI in order to explore their experience of participation. The verbatim transcripts were analyzed, and the end result was an overall theme that included 4 emerging categories. The theme "participation possibility" denotes the eventual involvement in life situations. Within this theme, there were 4 categories that emerged as a representation of essential aspects along the continuum of reclaiming participation: (1) dealing with the new self, (2) a journey dominated by obstacles, (3) the catalyst of participation, and (4) becoming an agent. Dealing with and recognizing these intervening conditions seem critical for clients to reconstruct the meaning that is necessary for a broader conception of participation. The transferability of these findings to dissimilar contexts may be limited. Client perspectives on reclaiming participation after injury affirm the notion of eventual participation, with each person finding strategies to succeed. In order to help clients reclaim participation, health professionals should develop contextually sensitive programs that include peer mentoring and reduce the influence of hindering factors.


Host-derived interleukin-18 differentially impacts regulatory and conventional T cell expansion during acute graft-versus-host disease.
Interleukin (IL)-18 stimulates T helper 1 (Th1) and Th2-mediated immune responses, and has been shown to modulate acute graft-versus-host disease (aGVHD). It is still unknown whether increased IL-18 levels during aGVHD are of host or donor origin, and how the absence of IL-18 has an impact on migration and expansion of conventional CD4(+)CD25(-)(Tconv) and CD4(+)CD25(+) regulatory (Treg) T cells in vivo. By utilizing IL-18 gene-deficient donor versus recipient animals we found that the major cytokine production during the early phase of aGVHD induction was recipient derived, whereas donor hematopoietic cells contributed significantly less. By generating IL-18(-/-) luciferase transgenic mice we were able to investigate the impact of IL-18 on Tconv and Treg expansion and trafficking with in vivo bioluminescence imaging. Although migration to secondary lymphoid organs did not have a significantly impact from the absence of host IL-18, Tconv but not Treg expansion increased significantly. Absence of host IL-18 production translated into lower IFN-gamma levels in the early phase after transplantation. We conclude that host-derived IL-18 is a major factor for IFN-gamma production that may have a protective effect on CD4(+)-mediated aGVHD, but is nonessential for Treg expansion in an allogeneic environment.


Abnormal brain tryptophan metabolism and clinical correlates in Tourette syndrome.
Symptoms in Tourette syndrome (TS) are likely related to abnormalities involving multiple neurotransmitter systems in striatal-thalamo-cortical circuitry. Although prior studies have found abnormal levels of tryptophan, serotonin, and their metabolites in blood, cerebrospinal fluid and brain tissue of TS patients, understanding of focal brain disturbances and their relationship to clinical phenotype remains poor. We used alpha-[(11)C]methyl-L-tryptophan (AMT) positron emission tomography (PET) to assess global and focal brain abnormalities of tryptophan metabolism and their relationship to behavioral phenotype in 26 children with TS and nine controls. Group comparisons on regional cortical and subcortical AMT uptake revealed decreased AMT uptake in bilateral dorsolateral prefrontal cortical and bilaterally increased uptake in the thalamus (P = 0.001) in TS children. The ratio of AMT uptake in subcortical structures to dorsolateral prefrontal cortex was significantly increased bilaterally (P < 0.01) in TS patients also. Behaviorally defined subgroups within the TS sample revealed differences in the pattern of AMT uptake in the fronto-striatal-thalamic circuit. This study demonstrates cortical and subcortical abnormalities of tryptophan metabolism in TS and provides neuroimaging evidence for a role of serotonergic mechanisms in the pathophysiology of TS.


Stability of dietary patterns assessed with reduced rank regression; the Zutphen Elderly Study.
Reduced rank regression (RRR) combines exploratory analysis with a-priori knowledge by including risk factors in the model. Dietary patterns, derived from RRR analysis, can be interpreted by the chosen risk factor profile and give an indication of positive or adverse health effects for a specific disease. Our aim was to assess the stability of dietary patterns derived by RRR over time. We used data from 467 men, aged 64-85 years, participating in the 1985 and 1990 examination rounds of the Zutphen Elderly Study. Backwards regression on risk factors and food groups was applied prior to the RRR analysis to exclude food groups with low predictability (from 36 to 19 food groups) for the chosen risk factor profile. For the final RRR analysis, dietary intake data from 19 food groups as predictor variables and 6 established risk factors for cardiovascular diseases (body mass index, systolic and diastolic blood pressure, high density lipoprotein and total cholesterol levels, and uric acid) were used. Three RRR dietary patterns were derived for both examination years: a "(low in) cereal fibre pattern", an "alcohol pattern" and an "inconsistent pattern". The "(low in) cereal fibre pattern" was most stable over time, with a correlation coefficient of 0.47 (95% CI: 0.38-0.53) between 1985 and 1990 measurements. Dietary patterns as measured by RRR, after backwards regression, are reasonably stable over a period of five years. Thus, RRR appears to be an attractive method to measure long-term dietary exposure for nutritional epidemiological studies, with one dietary measurement at baseline.


Heterochromatin drives compartmentalization of inverted and conventional nuclei.
The nucleus of mammalian cells displays a distinct spatial segregation of active euchromatic and inactive heterochromatic regions of the genome1,2. In conventional nuclei, microscopy shows that euchromatin is localized in the nuclear interior and heterochromatin at the nuclear periphery1,2. Genome-wide chromosome conformation capture (Hi-C) analyses show this segregation as a plaid pattern of contact enrichment within euchromatin and heterochromatin compartments3, and depletion between them. Many mechanisms for the formation of compartments have been proposed, such as attraction of heterochromatin to the nuclear lamina2,4, preferential attraction of similar chromatin to each other1,4-12, higher levels of chromatin mobility in active chromatin13-15 and transcription-related clustering of euchromatin16,17. However, these hypotheses have remained inconclusive, owing to the difficulty of disentangling intra-chromatin and chromatin-lamina interactions in conventional nuclei18. The marked reorganization of interphase chromosomes in the inverted nuclei of rods in nocturnal mammals19,20 provides an opportunity to elucidate the mechanisms that underlie spatial compartmentalization. Here we combine Hi-C analysis of inverted rod nuclei with microscopy and polymer simulations. We find that attractions between heterochromatic regions are crucial for establishing both compartmentalization and the concentric shells of pericentromeric heterochromatin, facultative heterochromatin and euchromatin in the inverted nucleus. When interactions between heterochromatin and the lamina are added, the same model recreates the conventional nuclear organization. In addition, our models allow us to rule out mechanisms of compartmentalization that involve strong euchromatin interactions. Together, our experiments and modelling suggest that attractions between heterochromatic regions are essential for the phase separation of the active and inactive genome in inverted and conventional nuclei, whereas interactions of the chromatin with the lamina are necessary to build the conventional architecture from these segregated phases.


Changes in Surface Free Energy and Surface Conductivity of Carbon Nanotube/Polyimide Nanocomposite Films Induced by UV Irradiation.
Changes in surface energy and electrical conductivity of polyimide (PI)-based nanocomposite films filled with carbon nanotubes (CNTs) induced by UV exposure are gaining considerable interest in microelectronic, aeronautical, and aerospace applications. However, the underlying mechanism of PI photochemistry and oxidation reactions induced by UV irradiation upon the surface in the presence of CNTs is still not clear. Here, we probed the interplay between CNTs and PIs under UV exposure in the surface properties of CNT/PI nanocomposite films. Changes in contact angles and surface electrical conductivity at the surface of CNT/PI nanocomposite films after UV exposure were measured. The unpaired electron intensity of free radicals generated by UV exposure was monitored by electron paramagnetic resonance. Our study indicates that the covalent interactions between CNTs and radicals generated by UV irradiation on the PI surfaces tailor the surface energy and surface conductivity through anchoring radicals on CNTs. Surprisingly, adding CNTs into PI films exposed to UV leads to antagonistic contributions of dispersion and polar components to the surface energy. The surface electrical conductivity of the CNT/PI nanocomposite films has been improved due to an enhanced hopping behavior with dense π-conjugated CNT sites. To explain the observed changes in surface energy and surface conductivity of CNT/PI nanocomposite films induced by UV exposure, a qualitative model was put forward describing the covalent interactions between UV-induced PI free radicals and CNTs, which govern the chemical nature of surface components. This study is helpful for characterizing and optimizing nanocomposite surface properties by tuning the covalent interactions between components at the nanoscale.


Analysis of the intronic single nucleotide polymorphism rs#466452 of the nephrin gene in patients with diabetic nephropathy.
We present the analysis of an intronic polymorphism of the nephrin gene and its relationship to the development of diabetic nephropathy in a study of diabetes type 1 and type 2 patients. The frequency of the single nucleotide polymorphism rs#466452 in the nephrin gene was determined in 231 patients and control subjects. The C/T status of the polymorphism was assessed using restriction enzyme digestions and the nephrin transcript from a kidney biopsy was examined. Association between the polymorphism and clinical parameters was evaluated using multivariate correspondence analysis. A bioinformatics analysis of the single nucleotide polymorphism rs#466452 suggested the appearance of a splicing enhancer sequence in intron 24 of the nephrin gene and a modification of proteins that bind to this sequence. However, no change in the splicing of a nephrin transcript from a renal biopsy was found. No association was found between the polymorphism and diabetes or degree of renal damage in diabetes type 1 or 2 patients. The single nucleotide polymorphism rs#466452 of the nephrin gene seems to be neutral in relation to diabetes and the development of diabetic nephropathy, and does not affect the splicing of a nephrin transcript, in spite of a splicing enhancer site.


An integrated genomic analysis of human glioblastoma multiforme.
Glioblastoma multiforme (GBM) is the most common and lethal type of brain cancer. To identify the genetic alterations in GBMs, we sequenced 20,661 protein coding genes, determined the presence of amplifications and deletions using high-density oligonucleotide arrays, and performed gene expression analyses using next-generation sequencing technologies in 22 human tumor samples. This comprehensive analysis led to the discovery of a variety of genes that were not known to be altered in GBMs. Most notably, we found recurrent mutations in the active site of isocitrate dehydrogenase 1 (IDH1) in 12% of GBM patients. Mutations in IDH1 occurred in a large fraction of young patients and in most patients with secondary GBMs and were associated with an increase in overall survival. These studies demonstrate the value of unbiased genomic analyses in the characterization of human brain cancer and identify a potentially useful genetic alteration for the classification and targeted therapy of GBMs.


Oncogenic Signaling Pathways in The Cancer Genome Atlas.
Genetic alterations in signaling pathways that control cell-cycle progression, apoptosis, and cell growth are common hallmarks of cancer, but the extent, mechanisms, and co-occurrence of alterations in these pathways differ between individual tumors and tumor types. Using mutations, copy-number changes, mRNA expression, gene fusions and DNA methylation in 9,125 tumors profiled by The Cancer Genome Atlas (TCGA), we analyzed the mechanisms and patterns of somatic alterations in ten canonical pathways: cell cycle, Hippo, Myc, Notch, Nrf2, PI-3-Kinase/Akt, RTK-RAS, TGFβ signaling, p53 and β-catenin/Wnt. We charted the detailed landscape of pathway alterations in 33 cancer types, stratified into 64 subtypes, and identified patterns of co-occurrence and mutual exclusivity. Eighty-nine percent of tumors had at least one driver alteration in these pathways, and 57% percent of tumors had at least one alteration potentially targetable by currently available drugs. Thirty percent of tumors had multiple targetable alterations, indicating opportunities for combination therapy.


Structure of the Rtt109-AcCoA/Vps75 complex and implications for chaperone-mediated histone acetylation.
Yeast Rtt109 promotes nucleosome assembly and genome stability by acetylating K9, K27, and K56 of histone H3 through interaction with either of two distinct histone chaperones, Vps75 or Asf1. We report the crystal structure of an Rtt109-AcCoA/Vps75 complex revealing an elongated Vps75 homodimer bound to two globular Rtt109 molecules to form a symmetrical holoenzyme with a ∼12 Å diameter central hole. Vps75 and Rtt109 residues that mediate complex formation in the crystals are also important for Rtt109-Vps75 interaction and H3K9/K27 acetylation both in vitro and in yeast cells. The same Rtt109 residues do not participate in Asf1-mediated Rtt109 acetylation in vitro or H3K56 acetylation in yeast cells, demonstrating that Asf1 and Vps75 dictate Rtt109 substrate specificity through distinct mechanisms. These studies also suggest that Vps75 binding stimulates Rtt109 catalytic activity by appropriately presenting the H3-H4 substrate within the central cavity of the holoenzyme to promote H3K9/K27 acetylation of new histones before deposition.


Effect of Restorative System and Thermal Cycling on the Tooth-Restoration Interface - OCT Evaluation.
The present study evaluated the tooth/noncarious cervical lesion restoration interface when using different adhesive systems and resin composites, submitted to thermal cycling (TC), using optical coherence tomography (OCT). Noncarious cervical lesion (NCCL) preparations (0.7 mm depth × 2 mm diameter) were performed on 60 human third molars and randomly divided into six groups, according to the adhesive system and resin composite used: group 1 = Adper Single Bond 2 (SB2) + Aelite LS Posterior (AP); group 2 = SB2 + Venus Diamond (VD); group = SB2 + Filtek Z250XT (Z250); group 4 = Clearfil SE Bond (CSE) + AP; group 5 = CSE + VD; group 6 = CSE + Z250. Selective enamel etching was performed for 30 seconds on groups 4, 5, and 6, while groups 1, 2, and 3 were etched for 30 seconds in enamel and 15 seconds in dentin. All groups were evaluated using OCT before and after TC (n=10). Images were analyzed using Image J software; enamel and dentin margins were separately evaluated. Data from OCT were submitted to PROC MIXED for repeated measurements and Tukey Kramer test (α = 0.05). No marginal gaps were observed in etched enamel, either before or after TC, for all adhesive and resin composite systems. A significant interaction was found between adhesive system and TC for the dentin groups; after TC, restorations with CSE showed smaller gaps at the dentin/restoration interface compared with SB2 for all resin composites. Increased gap percentages were noticed after TC compared with the gaps before TC for all groups. In conclusion, TC affected marginal integrity only in dentin margins, whereas etched enamel margins remained stable even after TC. Dentin margins restored with CSE adhesive system showed better marginal adaptation than those restored with SB2. Resin composites did not influence marginal integrity of NCCL restorations.


Impact of forest plantation on methane emissions from tropical peatland.
Tropical peatlands are a known source of methane (CH4 ) to the atmosphere, but their contribution to atmospheric CH4 is poorly constrained. Since the 1980s, extensive areas of the peatlands in Southeast Asia have experienced land-cover change to smallholder agriculture and forest plantations. This land-cover change generally involves lowering of groundwater level (GWL), as well as modification of vegetation type, both of which potentially influence CH4 emissions. We measured CH4 exchanges at the landscape scale using eddy covariance towers over two land-cover types in tropical peatland in Sumatra, Indonesia: (a) a natural forest and (b) an Acacia crassicarpa plantation. Annual CH4 exchanges over the natural forest (9.1 ± 0.9 g CH4 m-2 year-1 ) were around twice as high as those of the Acacia plantation (4.7 ± 1.5 g CH4 m-2 year-1 ). Results highlight that tropical peatlands are significant CH4 sources, and probably have a greater impact on global atmospheric CH4 concentrations than previously thought. Observations showed a clear diurnal variation in CH4 exchange over the natural forest where the GWL was higher than 40 cm below the ground surface. The diurnal variation in CH4 exchanges was strongly correlated with associated changes in the canopy conductance to water vapor, photosynthetic photon flux density, vapor pressure deficit, and air temperature. The absence of a comparable diurnal pattern in CH4 exchange over the Acacia plantation may be the result of the GWL being consistently below the root zone. Our results, which are among the first eddy covariance CH4 exchange data reported for any tropical peatland, should help to reduce the uncertainty in the estimation of CH4 emissions from a globally important ecosystem, provide a more complete estimate of the impact of land-cover change on tropical peat, and develop science-based peatland management practices that help to minimize greenhouse gas emissions.


Fallopian Tube Carcinoma.
Primary fallopian tube carcinoma is a rare and difficult to cure disease. It is often grouped under the epithelial ovarian cancer umbrella, together with primary ovarian and peritoneal carcinomas. More recent evidence has suggested that epithelial ovarian cancers originate from a fallopian tube precursor. The mainstay of treatment is surgical cytoreduction and platinum-based chemotherapy. There is much debate over the best timing for surgery and the best approach to delivering the chemotherapy: traditional intravenous once every 3 weeks regimen, versus intraperitoneal, versus dose-dense intravenous regimens. Although these debates continue, novel targeted therapies, including bevacizumab and poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP) inhibitors, have emerged. PARP inhibitors are particularly efficacious in patients with BRCA1/2 gene mutations, and their use has been shown to prolong patient survival. This article reviews the pathologic etiology; describes the heredity, treatment challenges, and controversies; and summarizes novel therapies in primary fallopian tube carcinoma.


The effects of nabilone on sleep in fibromyalgia: results of a randomized controlled trial.
Sleep disorders affect many patients with chronic pain conditions. Cannabis has been reported by several patient populations to help sleep. We evaluated the safety and efficacy of nabilone, a synthetic cannabinoid, on sleep disturbance in fibromyalgia (FM), a disease characterized by widespread chronic pain and insomnia. We conducted a randomized, double-blind, active-control, equivalency crossover trial to compare nabilone (0.5-1.0 mg before bedtime) to amitriptyline (10-20 mg before bedtime) in patients with FM with chronic insomnia. Subjects received each drug for 2 wk with a 2-wk washout period. The primary outcome was sleep quality, measured by the Insomnia Severity Index and the Leeds Sleep Evaluation Questionnaire. Secondary outcomes included pain, mood, quality of life, and adverse events (AEs). Thirty-one subjects were enrolled and 29 completed the trial (26 women, mean age 49.5 yr). Although sleep was improved by both amitriptyline and nabilone, nabilone was superior to amitriptyline (Insomnia Severity Index difference = 3.2; 95% confidence interval = 1.2-5.3). Nabilone was marginally better on the restfulness (Leeds Sleep Evaluation Questionnaire difference = 0.5 [0.0-1.0]) but not on wakefulness (difference = 0.3 [-0.2 to 0.8]). No effects on pain, mood, or quality of life were observed. AEs were mostly mild to moderate and were more frequent with nabilone. The most common AEs for nabilone were dizziness, nausea, and dry mouth. Nabilone is effective in improving sleep in patients with FM and is well tolerated. Low-dose nabilone given once daily at bedtime may be considered as an alternative to amitriptyline. Longer trials are needed to determine the duration of effect and to characterize long-term safety.


Peri-implant diseases: Consensus Report of the Sixth European Workshop on Periodontology.
Issues related to peri-implant disease were discussed. It was observed that the most common lesions that occur, i.e. peri-implant mucositis and peri-implantitis are caused by bacteria. While the lesion of peri-implant mucositis resides in the soft tissues, peri-implantitis also affects the supporting bone. Peri-implant mucositis occurs in about 80% of subjects (50% of sites) restored with implants, and peri-implantitis in between 28% and 56% of subjects (12-40% of sites). A number of risk indicators were identified including (i) poor oral hygiene, (ii) a history of periodontitis, (iii) diabetes and (iv) smoking. It was concluded that the treatment of peri-implant disease must include anti-infective measures. With respect to peri-implant mucositis, it appeared that non-surgical mechanical therapy caused the reduction in inflammation (bleeding on probing) but also that the adjunctive use of antimicrobial mouthrinses had a positive effect. It was agreed that the outcome of non-surgical treatment of peri-implantitis was unpredictable. The primary objective of surgical treatment in peri-implantitis is to get access to the implant surface for debridement and decontamination in order to achieve resolution of the inflammatory lesion. There was limited evidence that such treatment with the adjunctive use of systemic antibiotics could resolve a number of peri-implantitis lesions. There was no evidence that so-called regenerative procedures had additional beneficial effects on treatment outcome.


Advances in bone biology and new treatments for bone loss.
Recent advances in bone biology have led to a more detailed understanding of bone remodeling which is a process that leads to resorption of old bone and replacement by formation of new bone. The most important discoveries in this process of bone remodeling were those of the RANK Ligand/RANK/OPG system which is now recognized the dominant pathway regulating bone resorption. RANK Ligand (RANKL) is a cytokine belonging to the tumor necrosis factor family and is expressed by osteoblasts; it binds to membrane bound receptor RANK on osteoclasts and promotes differentiation of marrow cells through various stages to multinucleated osteoclasts which resorb bone. Several hormones such as parathyroid hormone, calcitriol and prostaglandins stimulate RANK Ligand expression by osteoblasts. Osteoblasts also secrete osteoprotegerin (OPG) which is a soluble receptor that is a potent antagonist of osteoclast formation by binding and inactivating RANKL and OPG is therefore an important regulator of bone resorption. OPG is stimulated by estrogen. OPG has been genetically engineered and in human subjects is a potent inhibitor of bone resorption. Another method for preventing bone resorption is to develope antibodies against RANKL and this has been shown to be a successful strategy. A single subcutaneous injection of this antibody (Denosumab) every 6 months proved to be a potent inhibitor of bone resorption and clinical fracture trials using this agent are now underway. These are novel developments that have risen from basic research in bone biology and other discoveries in the bone remodeling process can be expected to lead to further treatment options for various bone diseases.


Center-driven and Clinically Driven Variation in US Liver Transplant Maintenance Immunosuppression Therapy: A National Practice Patterns Analysis.
Variation in the use of immunosuppression regimens after liver transplant has not been well described. Immunosuppression regimens used after liver transplant were identified in a novel database integrating national transplant registry and pharmacy fill records for 24 238 recipients (2006-2014). Bilevel hierarchical models were developed to quantify the effects of transplant program, recipient, and donor characteristics on regimen choice. In the first 6 months after transplant, triple immunosuppression (tacrolimus, antimetabolite, corticosteroids) was the most common regimen (42.9%). By months 7 to 12, immunosuppression regimens were more commonly antimetabolite sparing (33.7%) or steroid sparing (26.9%), followed by triple (14.4%), mammalian target of rapamycin inhibitor (mTORi)-based (12.1%), or cyclosporine-based (9.2%). Based on intraclass correlation analysis, clinical characteristics explained less than 10% of the variation in immunosuppression choice, whereas program preference/practice explained 23% of steroid sparing, 26% of antimetabolite sparing, 28% of mTORi, and 21% of cyclosporine-based regimen use. Although case factors were not dominant practice drivers, triple immunosuppression in months 7 to 12 was more common among retransplant recipients and those with prior acute rejection. Hepatocellular carcinoma as cause of liver failure (adjusted odds ratio [aOR], 2.15; P<0.001), cancer within 6 months (aOR, 6.07; P<0.001), and 6-month estimated glomerular filtration rate less than 30 mL/min per 1.3 m2 (aOR, 1.98; P<0.001) were associated with mTORi use compared with triple immunosuppression in months 7 to 12, whereas acute rejection predicted lower use (aOR, 0.72; P=0.003). Liver transplant immunosuppression is dominantly driven by program preference, but case factors also affect regimen choice. This variation frames a natural experiment for future evaluations of comparative efficacy.


Increased susceptibility to repeated freeze-thaw cycles in Escherichia coli following long-term evolution in a benign environment.
In order to study the dynamics of evolutionary change, 12 populations of E. coli B were serially propagated for 20,000 generations in minimal glucose medium at constant 37 degrees C. Correlated changes in various other traits have been previously associated with the improvement in competitive fitness in the selective environment. This study examines whether these evolved lines changed in their ability to tolerate the stresses of prolonged freezing and repeated freeze-thaw cycles during adaptation to a benign environment. All 12 lines that evolved in the benign environment for 20,000 generations are more sensitive to freeze-thaw cycles than their ancestor. The evolved lines have an average mortality rate of 54% per daily cycle, compared to the ancestral rate of 34%. By contrast, there was no significant difference between the evolved lines and their ancestor in mortality during prolonged freezing. There was also some variability among the evolved lines in susceptibility to repeated freeze-thaw cycles. Those lines that had evolved higher competitive fitness in the minimal glucose medium at 37 degrees C also had higher mortality during freeze-thaw cycles. This variability was not associated, however, with differences among lines in DNA repair functionality and mutability. The consistency of the evolutionary declines in freeze-thaw tolerance, the correlation between fitness in glucose medium at 37 degrees C and mortality during freeze-thaw cycles, and the absence of greater declines in freeze-thaw survival among the hypermutable lines all indicate a trade-off between performance in minimal glucose medium at 37 degrees C and the capacity to tolerate this stress. Analyses of the mutations that enhance fitness at 37 degrees C may shed light on the physiological basis of this trade-off.


Urinary tract infections in a university hospital: pathogens and antibiotic susceptibility.
A substantial group of patients visit the emergency department (ED) with complaints of urinary tract infections (UTI). Treatment advice is based on national and local public health surveillance data. It is unclear whether this advice is adequate for hospitals with selected patient populations, such as university hospitals. We performed a retrospective study on patients visiting the ED of the Erasmus University Medical Center (Erasmus MC) in the Netherlands from January 1st, 2013 until December 31st, 2014 with a suspected complicated UTI (cUTI) and positive urinary cultures. Patient data, data concerning the ED visit and microbiological data were analysed. 439 patients visited the ED, of whom 429 had a cUTI. Our results were compared with NethMap data. Distribution of uropathogens was comparable with the overall distribution in the Netherlands. Antibiotic susceptibility was comparable for intravenous antibiotics, but was lower for oral antibiotics. Susceptibility for empiric antibiotic therapy (i.e., cefuroxime and gentamyicin) was 96.2%. Pathogens differed from the index culture in 56.2% 104/185) of the urinary cultures available from the previous year. Using logistic regression, we found that a shorter time between last admission to the initiated antibiotic regimen was associated with lower susceptibility of cultured uropathogens. The distribution and antibiotic susceptibility of uropathogens for intravenous antibiotics in a Dutch university hospital is comparable with overall distribution in the Netherlands. Empiric antibiotic therapy in our local guideline appears to be an adequate antibiotic regimen for cUTI and we therefore recommend treating patients accordingly. Extension of the chosen regimen based on earlier cultured pathogens is advised, and narrowing of the antibiotic regimen strongly discouraged.


Last and least: Findings on intrahousehold undernutrition from participatory research in South Asia.
We analyze intrahousehold mechanisms that rural South Asian women themselves identify as contributing to their inabilities to achieve adequate nutrition. The majority of our female respondents explain that, within their families, they are most likely to eat least and last and to seek lower quality healthcare later, both of which can lead to undernutrition. We analyze the explanations that women provide for why they do so. In addition to norm abidance, we highlight two social factors: a partner's heavy alcohol use and intimate partner violence. Women report that these factors adversely impact their own health and food security status and potentially contribute to differential nutritional outcomes within the same household. The emic perspectives presented here show us: (1) many rural South Asian women report shouldering a relatively greater share of food and nutrition insecurity within their households, and (2) taking their perspectives seriously assists in identifying the underlying contributors to intrahousehold differences. These findings indicate the value a gendered perspective on both the drivers of and measurement of undernutrition. Attending to these intrahousehold factors in future research may yield important insights for understanding drivers of nutritional outcomes.


Controlled expiration in mechanically-ventilated patients with chronic obstructive pulmonary disease (COPD).
In patients with severe chronic obstructive pulmonary disease (COPD), lung emptying may be affected by flow limitation. We tested the hypothesis that the airway compression leading to flow limitation can be counteracted by controlling the expiratory flow. The effects of an external resistor on lung emptying were studied in six patients with COPD, who were mechanically ventilated whilst sedated and paralysed. Respiratory mechanics were obtained during ventilatory support with and without the resistor. Airway compression was assessed using the interruptor method. For the study, a turbulent resistor was applied with the highest resistance level that did not increase the end-expiratory lung volume. At this resistance level, external positive end-expiratory pressure (PEEP) was generated in all patients. As total PEEP levels remained unchanged at both settings during the controlled expiration, the levels of intrinsic PEEP were significantly decreased from 0.96+/-0.30 to 0.53+/-0.19 kPa (mean+/-SD). Comparison of the expiratory flow-volume curves at both settings revealed that, during the controlled expiration, the flows were significantly decreased during the first 40% of the expired volume and significantly increased during the last 60%. As the end-expiratory lung volumes remained unchanged during both settings, these increments in flow indicated a decrease in effective resistance. Airway compression was observed during unimpeded expirations in all patients using the interruptor method. During the application of the resistor, airway compression was no longer detectable. In patients with chronic obstructive pulmonary disease receiving ventilatory support, the application of an external resistor could decrease effective expiratory resistance by counteracting airway compression, without increments in end-expiratory lung volume.


Fabrication of Janus Membranes for Desalination of Oil-Contaminated Saline Water.
Desalination of oil-contaminated saline water using membrane distillation requires hydrophobic membranes with underwater superoleophobic surfaces. For designing such membranes, the chemistry and morphology of the interfacial domains in contact with the contaminated water need to be adjusted such that a stable water layer, adhering to the surface, prevents oil droplets from wetting the membrane. In this article, we present an approach that relies on the controlled functionalization of the surface of polyvinylidene fluoride (PVDF) membranes; we adjust the surface topography of the membranes and introduce chemical heterogeneity to them. We show that the morphology of the PVDF surface can be altered by adjusting the composition of the nonsolvent bath used for the phase inversion process. Also, we render the surface of the membranes hydrophilic by using an alkaline chemical bath solution. The membrane morphology and effectiveness of our chemical treatment were confirmed by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), Fourier-transformed infrared spectroscopy (FTIR), and zeta potential measurements. A stable underwater contact angle, higher than 150°, was observed for both canola oil (ρ ≈ 0.913 g cm-3, γ ≈ 31.5 mN m-1) and hexane (ρ ≈ 0.655 g cm-3, γ ≈ 18 mN m-1). We evaluated the performance of both pristine and functionalized membranes in a laboratory-scale direct contact membrane distillation (DCMD) setup and desalinated a saline solution contaminated with 500 ppm canola oil. Our results show that oil does not wet the functionalized membrane during the desalination process. The average permeate flux and salt rejection values for the functionalized membranes were 45 ± 5 Lm-2h-1 ( Tfeed = 70 °C, Tdistillate = 20 °C) and 99.99%, respectively.


Demyelination and remyelination after spinal cord injury.
Since the discovery in the 1960s that remyelination can occur in the damaged central nervous system (CNS) (Bunge et al. 1961), there has been much progress in understanding the cellular and molecular biology of oligodendroglia and the factors that regulate their propagation, migration, differentiation, maturation, and ability to myelinate nerve axons. More recently, greater understanding of disease states and the role of oligodendrocytes in remyelination have sparked tremendous interest in this once obscure field. Although the explosion of information is being hampered by adherence to commonly held beliefs based on empirical evidence, novel molecular and cellular tools are allowing scientists to address age-old assumptions. It is now recognized that, as well as promoting salutatory conduction along axons, oligodendroglia are important near-term clinical targets for restoring function after CNS injury, particularly spinal cord injury. Thus, remyelination appears to be one of the most feasible restoration strategies. This review focuses on concepts that are important for developing strategies of repair. The brightest young scientists will be attracted into this exciting field by its near-term potential for human application.


Morphological consequences of artificial cranial deformation: Modularity and integration.
The cranium is an anatomically complex structure. One source of its complexity is due to its modular organization. Cranial modules are distinct and partially independent units that interact substantially during ontogeny thus generating morphological integration. Artificial Cranial Deformation (ACD) occurs when the human skull is intentionally deformed, through the use of different deforming devices applied to the head while it is developing. Hence, ACD provides an interesting example to assess the degree to which biomechanical perturbations of the developing neurocranium impact on the degree of morphological integration in the skull as a whole. The main objective of this study was to assess how ACD affects the morphological integration of the skull. This was accomplished by comparing a sample of non-deformed crania and two sets of deformed crania (i.e. antero-posterior and oblique). Both developmental and static modularity and integration were assessed through Generalized Procrustes Analysis by considering the symmetric and asymmetric components of variation in adults, using 3D landmark coordinates as raw data. The presence of two developmental modules (i.e. viscero and neurocranium) in the skull was tested. Then, in order to understand how ACD affects morphological integration, the covariation pattern between the neuro and viscerocranium was examined in antero-posterior, oblique and non-deformed cranial categories using Partial Least-Squares. The main objective of this study was to assess how ACD affects the morphological integration of the skull. This was accomplished by comparing a sample of deformed (i.e. antero-posterior and oblique) and non-deformed crania. Hence, differences in integration patterns were compared between groups. The obtained results support the modular organization of the human skull in the two analyzed modules. The integration analyses show that the oblique ACD style differentially affects the static morphological integration of the skull by increasing the covariance between neuro and viscerocranium in a more constrained way than in antero-posterior and non-deformed skulls. In addition, the antero-posterior ACD style seems to affect the developmental integration of the skull by directing the covariation pattern in a more defined manner as compared to the other cranial categories.


Effects of perindopril-indapamide on left ventricular diastolic function and mass in patients with type 2 diabetes: the ADVANCE Echocardiography Substudy.
The Action in Diabetes and Vascular Disease (ADVANCE) Study demonstrated that a fixed combination of perindopril and indapamide reduced the risk of major vascular events and mortality in patients with type 2 diabetes. This Echocardiographic Substudy was designed to determine the effects of this treatment on left ventricular diastolic function and left ventricular mass. Five hundred and fifty-five patients entering ADVANCE underwent quantitative echocardiography prior to randomization and after 6 months and 4 years of treatment with perindopril-indapamide or placebo. Main end points were left ventricular diastolic function (ratio of mitral E velocity/early medial mitral annular tissue Doppler velocity, E/Em, and left atrial volume index) and left ventricular mass index. Overall, blood pressure was reduced in the perindopril-indapamide group compared with placebo. E/Em and left atrial volume index both increased over the 4 years. There was no effect of perindopril-indapamide on E/Em, although there was a small attenuation of the increase in left atrial volume index with active treatment. Left ventricular mass index was reduced by 2.7 g/m with active treatment (95% confidence interval -5.0 to -0.1, P = 0.04). Compared with placebo, the perindopril-indapamide combination reduced blood pressure and left ventricular mass in patients with diabetes, but did not improve left ventricular diastolic function. Left ventricular diastolic function worsened in both groups over 4 years, despite blood pressure reduction and reduction in left ventricular mass. Improving left ventricular diastolic function remains a challenge in patients with diabetes.


Patterns of mortality in southern sea otters (Enhydra lutris nereis) from 1998-2001.
Detailed postmortem examination of southern sea otters (Enhydra lutris nereis) found along the California (USA) coast has provided an exceptional opportunity to understand factors influencing survival in this threatened marine mammal species. In order to evaluate recent trends in causes of mortality, the demographic and geographic distribution of causes of death in freshly deceased beachcast sea otters necropsied from 1998-2001 were evaluated. Protozoal encephalitis, acanthocephalan-related disease, shark attack, and cardiac disease were identified as common causes of death in sea otters examined. While infection with acanthocephalan parasites was more likely to cause death in juvenile otters, Toxoplasma gondii encephalitis, shark attack, and cardiac disease were more common in prime-aged adult otters. Cardiac disease is a newly recognized cause of mortality in sea otters and T. gondii encephalitis was significantly associated with this condition. Otters with fatal shark bites were over three times more likely to have pre-existing T. gondii encephalitis suggesting that shark attack, which is a long-recognized source of mortality in otters, may be coupled with a recently recognized disease in otters. Spatial clusters of cause-specific mortality were detected for T. gondii encephalitis (in Estero Bay), acanthocephalan peritonitis (in southern Monterey Bay), and shark attack (from Santa Cruz to Point Año Nuevo). Diseases caused by parasites, bacteria, or fungi and diseases without a specified etiology were the primary cause of death in 63.8% of otters examined. Parasitic disease alone caused death in 38.1% of otters examined. This pattern of mortality, observed predominantly in juvenile and prime-aged adult southern sea otters, has negative implications for the overall health and recovery of this population.


Does physical therapy still have a place in the treatment of ankylosing spondylitis?
To review studies of various physical therapy programs in ankylosing spondylitis and identify their benefits and potential indications in the treatment of this disease. Various exercise and physical therapy programs have been evaluated in clinical studies. Home exercise programs have been shown to improve symptoms, mobility, function and overall quality of life. Formal physical therapy under the supervision of a physical therapist has been shown to improve posture, fitness, mobility, function and mood. Water therapy may improve symptoms, function and overall sense of health. Inpatient rehabilitation may provide rapid short-term improvement in pain and stiffness, mobility, function and quality of life for patients with severe active disease. Despite the advances in the pharmacological therapy of ankylosing spondylitis, physical therapy remains an essential part of the management plan. Even though data are not sufficient to determine which specific physical therapy program should be recommended, physicians should implement such nonpharmacological therapy as part of a comprehensive management strategy for this disease. All patients should receive instructions on proper posture and home exercises and be encouraged to perform water exercises if they can. Formal physical therapy and, in most severe cases, inpatient rehabilitation may be of benefit to select patients with ankylosing spondylitis.


Modeling and design of irregularly arrayed waveguide gratings.
In this paper we propose a concept of irregularly arrayed waveguide gratings (IAWGs). By replacing regularly arrayed waveguides of conventional AWGs with irregularly arrayed ones, we found theoretically that the IAWGs have arbitrary free spectral ranges and are suitable to provide a large number of channels with relatively smaller circuit regions. A Fourier optics model is presented, which is able to calculate the transmission characteristics between any arbitrary pair of input/output ports in either conventional AWGs or IAWGs. As an example, a 1080-channel IAWG with 0.199 nm spacing is designed and simulated. Results show this device can be contained in a 6-inch wafer and provide two work patterns: all 1080 channels with a relatively high crosstalk of about -20 dB, and 128 successive channels which can be arbitrarily moveable over the range of the 1080 channels with a crosstalk of about -32 dB.


Helping patients decide: ten steps to better risk communication.
With increasing frequency, patients are being asked to make complex decisions about cancer screening, prevention, and treatment. These decisions are fraught with emotion and cognitive difficulty simultaneously. Many Americans have low numeracy skills making the cognitive demands even greater whenever, as is often the case, patients are presented with risk statistics and asked to make comparisons between the risks and benefits of multiple options and to make informed medical decisions. In this commentary, we highlight 10 methods that have been empirically shown to improve patients' understanding of risk and benefit information and/or their decision making. The methods range from presenting absolute risks using frequencies (rather than presenting relative risks) to using a risk format that clarifies how treatment changes risks from preexisting baseline levels to using plain language. We then provide recommendations for how health-care providers and health educators can best to communicate this complex medical information to patients, including using plain language, pictographs, and absolute risks instead of relative risks.


Breast Cancer Risk After Recent Childbirth: A Pooled Analysis of 15 Prospective Studies.
Parity is widely recognized as protective for breast cancer, but breast cancer risk may be increased shortly after childbirth. Whether this risk varies with breastfeeding, family history of breast cancer, or specific tumor subtype has rarely been evaluated. To characterize breast cancer risk in relation to recent childbirth. Pooled analysis of individual-level data from 15 prospective cohort studies. The international Premenopausal Breast Cancer Collaborative Group. Women younger than 55 years. During 9.6 million person-years of follow-up, 18 826 incident cases of breast cancer were diagnosed. Hazard ratios (HRs) and 95% CIs for breast cancer were calculated using Cox proportional hazards regression. Compared with nulliparous women, parous women had an HR for breast cancer that peaked about 5 years after birth (HR, 1.80 [95% CI, 1.63 to 1.99]) before decreasing to 0.77 (CI, 0.67 to 0.88) after 34 years. The association crossed over from positive to negative about 24 years after birth. The overall pattern was driven by estrogen receptor (ER)-positive breast cancer; no crossover was seen for ER-negative cancer. Increases in breast cancer risk after childbirth were pronounced when combined with a family history of breast cancer and were greater for women who were older at first birth or who had more births. Breastfeeding did not modify overall risk patterns. Breast cancer diagnoses during pregnancy were not uniformly distinguishable from early postpartum diagnoses. Data on human epidermal growth factor receptor 2 (HER2) oncogene overexpression were limited. Compared with nulliparous women, parous women have an increased risk for breast cancer for more than 20 years after childbirth. Health care providers should consider recent childbirth a risk factor for breast cancer in young women. The Avon Foundation, the National Institute of Environmental Health Sciences, Breast Cancer Now and the UK National Health Service, and the Institute of Cancer Research.


Richness and species composition of arboreal arthropods affected by nutrients and predators: a press experiment.
A longstanding goal for ecologists is to understand the processes that maintain biological diversity in communities, yet few studies have investigated the combined effects of predators and resources on biodiversity in natural ecosystems. We fertilized nutrient limited plots and excluded insectivorous birds in a randomized block design, and examined the impacts on arthropods associated with the dominant tree in the Hawaiian Islands, Metrosideros polymorpha (Myrtaceae). After 33 months, the species load (per foliage mass) of herbivores and carnivores increased with fertilization, but rarified richness (standardized to abundance) did not change. Fertilization depressed species richness of arboreal detritivores, and carnivore richness dropped in caged, unfertilized plots, both because of the increased dominance of common, introduced species with treatments. Herbivore species abundance distributions were more equitable than other trophic levels following treatments, and fertilization added specialized native species without changing relativized species richness. Overall, bird removal and nutrient addition treatments on arthropod richness acted largely independently, but with countervailing influences that obscured distinct top-down and bottom-up effects on different trophic levels. This study demonstrates that species composition, biological invasions, and the individuality of species traits may complicate efforts to predict the interactive effects of resources and predation on species diversity in food webs.


Laparoscopic and open surgical nephrectomy for xanthogranulomatous pyelonephritis.
Xanthogranulomatous pyelonephritis (XGP) is a severe, chronic renal-parenchymal infection. Nephrectomy is the treatment of choice. Because of the renal and perirenal inflammatory changes that commonly accompany XGP, the laparoscopic approach is difficult. We compared our experience with laparoscopic and open surgical nephrectomy for XGP. A retrospective chart review of all adult nephrectomy specimens with the pathologic diagnosis of XGP between January 1997 and May 2003 was performed. Preoperative presentation, operative details, and postoperative recovery and complications were included in the data collection. Three patients approached laparoscopically and eight patients approached with open surgery were found to have XGP on pathologic analysis. The disease was suspected preoperatively in all patients. Among the laparoscopically treated patients, there was 1 (33%) who suffered major complications; this was the only patient who required conversion to open surgery. Among the open-surgical group, there were 2 (22%) major and 3 (33%) minor complications. Postoperative hospitalization was longer in the open-surgical group (mean 13.7 v 4.7 days), and when the case of open conversion was excluded, narcotic use was less in the laparoscopy group. The treatment of some XGP cases with laparoscopic nephrectomy is a possible, albeit challenging, option. The incidences of intraoperative and postoperative complications were roughly equivalent in the laparoscopic and open-surgery patients in our study. If completed, laparoscopy appears to be associated with decreased postoperative morbidity. However, this may represent selection bias, and larger, prospective studies may better define the suspected benefit.


Comparison of horizontal and vertical dynamic visual acuity in patients with vestibular dysfunction and nonvestibular dizziness.
Blurred vision with head movement is a common symptom reported by patients with vestibular dysfunction affecting the vestibulo-ocular reflex (VOR). Impaired VOR can be measured by comparing visual acuity in which there is no head movement to visual acuity obtained with head movement. A previous study demonstrated that dynamic visual acuity (DVA) testing using vertical head movement revealed deficits in impaired VOR. There is evidence that horizontal head movement is more sensitive to impaired VOR. The objective of this investigation was to compare horizontal and vertical DVA in participants with normal vestibular function (NVF), impaired vestibular function (IVF), and participants with nonvestibular dizziness (NVD). Participants performed the visual acuity task in a baseline condition with no movement and also in two dynamic conditions, horizontal head movement and vertical head movement. Horizontal DVA was twice as sensitive to impaired VOR than vertical DVA. Results suggest that horizontal volitional head movement should be incorporated into tasks measuring functional deficits of impaired VOR.


Epidermal Growth Factor Receptor in Prostate Cancer Derived Exosomes.
Exosomes proteins and microRNAs have gained much attention as diagnostic tools and biomarker potential in various malignancies including prostate cancer (PCa). However, the role of exosomes and membrane-associated receptors, particularly epidermal growth factor receptor (EGFR) as mediators of cell proliferation and invasion in PCa progression remains unexplored. EGFR is frequently overexpressed and has been associated with aggressive forms of PCa. While PCa cells and tissues express EGFR, it is unknown whether exosomes derived from PCa cells or PCa patient serum contains EGFR. The aim of this study was to detect and characterize EGFR in exosomes derived from PCa cells, LNCaP xenograft and PCa patient serum. Exosomes were isolated from conditioned media of different PCa cell lines; LNCaP xenograft serum as well as patient plasma/serum by differential centrifugation and ultracentrifugation on a sucrose density gradient. Exosomes were confirmed by electron microscopy, expression of exosomal markers and NanoSight™ analysis. EGFR expression was determined by western blot analysis and ELISA. This study demonstrates that exosomes may easily be derived from PCa cell lines, serum obtained from PCa xenograft bearing mice and clinical samples derived from PCa patients. Presence of exosomal EGFR in PCa patient exosomes may present a novel approach for measuring of the disease state. Our work will allow to build on this finding for future understanding of PCa exosomes and their potential role in PCa progression and as minimal invasive biomarkers for PCa.


Rai1 duplication causes physical and behavioral phenotypes in a mouse model of dup(17)(p11.2p11.2).
Genomic disorders are conditions that result from DNA rearrangements, such as deletions or duplications. The identification of the dosage-sensitive gene(s) within the rearranged genomic interval is important for the elucidation of genes responsible for complex neurobehavioral phenotypes. Smith-Magenis syndrome is associated with a 3.7-Mb deletion in 17p11.2, and its clinical presentation is caused by retinoic acid inducible 1 (RAI1) haploinsufficiency. The reciprocal microduplication syndrome, dup(17)(p11.2p11.2), manifests several neurobehavioral abnormalities, but the responsible dosage-sensitive gene(s) remain undefined. We previously generated a mouse model for dup(17)(p11.2p11.2), Dp(11)17/+, that recapitulated most of the phenotypes observed in human patients. We have now analyzed compound heterozygous mice carrying a duplication [Dp(11)17] in one chromosome 11 along with a null allele of Rai1 in the other chromosome 11 homologue [Dp(11)17/Rai1(-) mice] in order to study the relationship between Rai1 gene copy number and the Dp(11)17/+ phenotypes. Normal disomic Rai1 gene dosage was sufficient to rescue the complex physical and behavioral phenotypes observed in Dp(11)17/+ mice, despite altered trisomic copy number of the other 18 genes present in the rearranged genomic interval. These data provide a model for variation in copy number of single genes that could influence common traits such as obesity and behavior.


Reciprocal regulation of myocardial microRNAs and messenger RNA in human cardiomyopathy and reversal of the microRNA signature by biomechanical support.
Much has been learned about transcriptional control of cardiac gene expression in clinical and experimental congestive heart failure (CHF), but less is known about dynamic regulation of microRNAs (miRs) in CHF and during CHF treatment. We performed comprehensive microarray profiling of miRs and messenger RNAs (mRNAs) in myocardial specimens from human CHF with (n=10) or without (n=17) biomechanical support from left ventricular assist devices in comparison to nonfailing hearts (n=11). Twenty-eight miRs were upregulated >2.0-fold (P<0.001) in CHF, with nearly complete normalization of the heart failure miR signature by left ventricular assist device treatment. In contrast, of 444 mRNAs that were altered by >1.3-fold in failing hearts, only 29 mRNAs normalized by as much as 25% in post-left ventricular assist device hearts. Unsupervised hierarchical clustering of upregulated miRs and mRNAs with nearest centroid analysis and leave-1-out cross-validation revealed that combining the miR and mRNA signatures increased the ability of RNA profiling to serve as a clinical biomarker of diagnostic group and functional class. These results show that miRs are more sensitive than mRNAs to the acute functional status of end-stage heart failure, consistent with important functions for regulated miRs in the myocardial response to stress. Combined miR and mRNA profiling may have superior potential as a diagnostic and prognostic test in end-stage cardiomyopathy.


Pump-Flow-Probe X-Ray Absorption Spectroscopy as a Tool for Studying Intermediate States of Photocatalytic Systems.
A new setup for pump-flow-probe X-ray absorption spectroscopy has been implemented at the SuperXAS beamline of the Swiss Light Source. It allows recording X-ray absorption spectra with a time resolution of tens of microseconds and high detection efficiency for samples with sub-mM concentrations. A continuous wave laser is used for the photoexcitation, with the distance between laser and X-ray beams and velocity of liquid flow determining the time delay, while the focusing of both beams and the flow speed define the time resolution. This method is compared with the alternative measurement technique that utilizes a 1 kHz repetition rate laser and multiple X-ray probe pulses. Such an experiment was performed at beamline 11ID-D of the Advanced Photon Source. Advantages, limitations and potential for improvement of the pump-flow-probe setup are discussed by analyzing the photon statistics. Both methods, with Co K-edge probing were applied to the investigation of a cobaloxime-based photo-catalytic reaction. The interplay between optimizing for efficient photoexcitation and time resolution as well as the effect of sample degradation for these two setups are discussed.


Poor sleep quality and erectile dysfunction in students from a Peruvian University: A cross-sectional study.
We aimed to evaluate the association between sleep quality and erectile dysfunction in young university students. A cross-sectional survey was conducted in men aged 18-30 years from Universidad Ricardo Palma, Lima, Peru. The survey comprised the International Index of Erectile Function, Pittsburgh Sleep Quality Index, Berlin questionnaire, and questions related to sociodemographic data. Prevalence ratios were estimated with generalized linear models. Of 381 participants, the median age was 23 years. Half of the students (50.9%) had poor sleep quality, of which 72.7% had mild erectile dysfunction and 20.6% mild to moderate dysfunction. Prevalence of erectile dysfunction was significantly higher in students with poor sleep quality than in students with good sleep quality (aPR = 6.48; 95% CI: 4.58-9.17) after adjusting for age, academic year, nutritional status, and sleep apnea. In a subsequent exploratory analysis, sleep apnea was associated with a higher prevalence of erectile dysfunction (aPR = 1.19; 95% CI: 1.01-1.39), while overweight (aPR = 0.85; 95% CI: 0.76-0.95) and obesity (aPR = 0.65; 95% CI: 0.52-0.82) were associated with a lower prevalence of this condition. Poor sleep quality was independently associated with erectile dysfunction in young university students. This finding suggests that male students are at risk for sexual problems due to possible academic demands and relationship issues.


Endogenous Wnt signaling maintains neural progenitor cell potency.
Wnt signaling regulates neural stem cell (NSC) function and development throughout an individual's lifetime. Intriguingly, adult hippocampal progenitors (AHPs) produce several Wnts, and the intracellular machinery necessary to respond to them, creating the potential for an active autocrine-signaling loop within this stem cell niche. However, the standard luciferase-based Wnt assay failed to detect this signaling loop. This assay is inherently less temporally sensitive to activity among a population of unsynchronized proliferating cells because it relies on the rapidly degrading reporter luciferase. We circumvented this limitation using a promoter assay that employs green fluorescent protein (GFP), as a relatively long-lived reporter of canonical Wnt activity. We found that at baseline, AHPs secreted functional Wnt that self-stimulates low-level canonical Wnt signaling. Elimination baseline Wnt activity, via application of an extracellular Wnt antagonist promoted neurogenesis, based on a combination of unbiased gene expression analysis and cell-fate analysis. A detailed clonal analysis of progenitors transduced with specific intracellular antagonists of canonical signaling, either Axin or truncated cadherin (beta-catenin sequestering), revealed that loss of baseline signaling depletes the population of multipotent precursors, thereby driving an increasing fraction to assume a committed cell fate (i.e., unipotent progenitors). Similarly, baseline Wnt signaling repressed differentiation of human NSCs. Although the specific Wnts produced by neural precursors vary with age and between species, their effects remain remarkably consistent. In sum, this study establishes that autonomous Wnt signaling is a conserved feature of the neurogenic niche that preserves the delicate balance between NSC maintenance and differentiation.


Beneficial effects of hydrogen gas against spinal cord ischemia-reperfusion injury in rabbits.
Recently, hydrogen gas (H₂) is reported to be a new therapeutic agent in organ damage induced by ischemia-reperfusion (I/R). The present study was designed to investigate the beneficial effects of H₂ against spinal cord I/R injury and its associated mechanisms. Spinal cord ischemia was induced by infrarenal aortic occlusion for 20 min in male New Zealand white rabbits. Treatment with 1%, 2% or 4% H₂ inhalation was given from 10 min before reperfusion to 60 min after reperfusion (total 70 min). Here, we found that I/R-challenged animals showed significant spinal cord damage characterized by the decreased numbers of normal motor neurons and hind-limb motor dysfunction, which was significantly improved by 2% and 4 % H₂ treatment. Furthermore, we found that the beneficial effects of H₂ treatment against spinal cord I/R injury were associated with the decreased levels of oxidative products [8-iso-prostaglandin F2α (8-iso-PGF2α) and malondialdehyde (MDA)] and pro-inflammatory cytokines [tumor necrosis factor-alpha (TNF-α) and high-mobility group box 1 (HMGB1)], as well as increased activities of antioxidant enzymes [superoxide dismutase (SOD) and catalase (CAT)] in serum and spinal cord. In addition, H₂ treatment reduced motor neuron apoptosis in the spinal cord of this model. Thus, H₂ inhalation may be an effective therapeutic strategy for spinal cord I/R damage.


Antidepressant use in Denmark, Germany, Spain, and Sweden between 2009 and 2014: Incidence and comorbidities of antidepressant initiators.
We aimed to describe patterns of use and characteristics of 10 commonly used antidepressants for the period 2009-2014 in Denmark, Germany, Spain, and Sweden. Adult initiators from 2009 to 2014 of each study antidepressant were identified in four countries using five data sources: the Danish National registers, GePaRD (Germany), EpiChron (Aragon, Spain), SIDIAP (Catalonia, Spain), and the Swedish National Registers. The study included 10 study antidepressants: citalopram, escitalopram, fluoxetine, paroxetine, sertraline, duloxetine, venlafaxine, amitriptyline, mirtazapine, and agomelatine. Citalopram was the most prescribed study antidepressant, followed by mirtazapine. Paroxetine and agomelatine were the least prescribed antidepressants. Mirtazapine was widely used among older antidepressant initiators with higher percentages of comorbidities at baseline, and fluoxetine was used among young patients. Citalopram and amitriptyline had the lowest percentage of multiple antidepressant use in the 12 months prior to the current treatment episode, while agomelatine, duloxetine, and venlafaxine had the highest percentage of multiple antidepressant use in the year prior to the current treatment episode. The most important limitations are exposure information based on filled prescriptions, focus on antidepressant initiators only, lack of information on the indication, and heterogeneity of the type of data across data sources. Results of this study including 4.8 million study antidepressant initiators of study antidepressants suggest that citalopram and mirtazapine are the most commonly prescribed antidepressants. Agomelatine and paroxetine were the least used antidepressants in the participating populations. Mirtazapine was the antidepressant most commonly prescribed among older antidepressant initiators with high percentage of comorbidities at baseline, whereas fluoxetine was commonly used among young patients.


A review of factors affecting the transfer of sexual and reproductive health training into practice in low and lower-middle income country humanitarian settings.
A lack of access to sexual and reproductive health (SRH) care is the leading cause of morbidity and mortality among displaced women and girls of reproductive age. Efforts to address this public health emergency in humanitarian settings have included the widespread delivery of training programmes to address gaps in health worker capacity for SRH. There remains a lack of data on the factors which may affect the ability of health workers to apply SRH knowledge and skills gained through training programmes in humanitarian contexts. We searched four electronic databases and ten key organizations' websites to locate literature on SRH training for humanitarian settings in low and lower-middle income countries. Papers were examined using content analysis to identify factors which contribute to health workers' capacity to transfer SRH knowledge, skills and attitudes learned in training into practice in humanitarian settings. Seven studies were included in this review. Six research papers focused on the response stage of humanitarian crises and five papers featured the disaster context of conflict. A range of SRH components were addressed including maternal, newborn health and sexual violence. The review identified factors, including appropriate resourcing, organisational support and confidence in health care workers that were found to facilitate the transfer of learning. The findings suggest the presence of factors that moderate the transfer of training at the individual, training, organisational, socio-cultural, political and health system levels. Supportive strategies are necessary to best assist trainees to apply newly acquired knowledge and skills in their work settings. These interventions must address factors that moderate the success of learning transfer. Findings from this review suggest that these are related to the individual trainee, the training program itself and the workplace as well as the broader environmental context. Organisations which provide SRH training for humanitarian emergencies should work to identify the system of moderating factors that affect training transfer in their setting and employ evidence-based strategies to ameliorate these.


The relationship between baseline nutritional status with subsequent parenteral nutrition and clinical outcomes in cancer patients undergoing hyperthermic intraperitoneal chemotherapy.
The combination of cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) is a promising treatment option for selected patients with peritoneal carcinomatosis. This retrospective study investigated the relationship between baseline nutritional assessment with subsequent parenteral nutritional (PN) and clinical outcomes in cancer patients undergoing CRS and HIPEC. A consecutive series of 60 patients undergoing CRS and HIPEC at our institution between January 2009 and May 2011. Subjective Global Assessment (SGA) was used to assess nutritional status. Patients were classified preoperatively as: well nourished (SGA-A), mildly-moderately malnourished (SGA-B), and severely malnourished (SGA-C). For PN, patients were divided into 2 groups: those who received PN (PN+) and those who did not receive PN (PN-). The primary outcomes of interest were length of stay (LOS), postoperative complications, ECOG performance status (PS) and survival. LOS was calculated as the number of days in the hospital post surgery. Performance status was measured on a scale of 0-4. Survival was calculated from the date of first visit to the date of death/last contact. Of 60 patients, 19 were males and 41 females. The mean age at presentation was 50.3 years. The most common cancer types were colorectal (n = 24) and gynecologic (n = 19) with the majority of patients (n = 47) treated previously before coming to our institution. 33 patients were SGA-A, 22 SGA-B and 5 SGA-C prior to surgery. Of a total of 60 patients, 31 received PN. Mean LOS for the entire cohort was 16.2 days (SD = 9.8). Mean LOS for preoperative SGA-A, SGA-B and SGA-C were 15.0, 15.2 and 27.8 days respectively (ANOVA p = 0.02). Overall incidence of complications was 26.7% (16/60). Complications were recorded in 9 of 33 (27.3%) preoperative SGA-A patients and 7 of 27 (25.9%) SGA-B + C patients (p = 0.91). The median overall survival was 17.5 months (95% CI = 13.0 to 22.1 months). Median survival for preoperative SGA-A and SGA-B + C cohorts was 22.4 and 10.4 months respectively (p = 0.006). The preoperative SGA predicts LOS and survival in cancer patients undergoing HIPEC. Future randomized clinical trials in this patient population should investigate the systematic provision of PN to all malnourished patients in the preoperative period for a minimum of 7-10 days with the continuation of PN in the postoperative period.


Inter-relationships between type, size and colour of fruits and dispersal in southern African trees.
The indigenous angiosperm tree flora (1,340 species) of southern Africa was analysed for type, size and colour of fruits and class of biotic dispersal agent (consumer). Species producing fleshy (drupes and berries) and dry (pods, capsules and nuts) fruits account for 52% and 47%, respectively, of the flora. The flora contains about 2.5 times as many berry-producing as drupe-producing species. Based on a log-linear model, fruit type, consumer and fruit size are dependent statistically on each other, whereas fruit colour depends on both fruit size and consumer type acting independently of each other. Drupes and berries are consumed by birds and mammals, with berries being favoured by both birds and mammals. At least 23% (307 species) of the flora apparently depends predominantly on birds for seed dispersal. Drupes and berries favoured by birds tend to be small and brightly coloured (red or black), whereas those eaten mainly by mammals tend to be large and dull (yellow or green). Relatively few fleshy fruits are brown. Pods, capsules and nuts tend to be brown or green. Birds apparently tend to avoid eating green fruits. The notion that green coloration has evolved to enhance crypsis and/or to signal unpalatability in unripe fruit to reduce premature exploitation is questioned. Green as a cryptic colour is incompatible with the demonstrated mammalian selection of this colour, while to function aposematically a stronger contrast colour may be required.


Periodic bilayer organization in the complexes of Beta-2 Glycoprotein I with anionic lipid membranes.
β2 glycoprotein I (β2GPI) is a soluble protein that participates in blood coagulation, clearance of apoptotic bodies and generation of antigens in antiphospholipid syndrome among many other functions. We studied the aggregates formed by β2GPI with the anionic phospholipids palmitoyloleoylphosphatidyl glycerol, dimyristoylphosphatidyl glycerol, dipalmitoylphosphatidyl glycerol and cardiolipin using small angle X-ray scattering. The complexes obtained in a medium containing 0.01 M NaCl showed Bragg peaks up to the sixth order in a well-defined integer sequence indicating the presence of a lamellar stacking with a periodicity of 17.8 nm and with largely reduced membrane fluctuations. Modeling the complex signal allowed us to conclude that the coherence length was only two bilayers and that about 15% of the total surface was actually stacked. The space between bilayers allows accommodating an extended β2GPI molecule making a bridge between the interacting bilayers. The interactions between membranes mediated by β2GPI was favored when the membranes were in the liquid crystalline state.


Mindfulness practice leads to increases in regional brain gray matter density.
Therapeutic interventions that incorporate training in mindfulness meditation have become increasingly popular, but to date little is known about neural mechanisms associated with these interventions. Mindfulness-Based Stress Reduction (MBSR), one of the most widely used mindfulness training programs, has been reported to produce positive effects on psychological well-being and to ameliorate symptoms of a number of disorders. Here, we report a controlled longitudinal study to investigate pre-post changes in brain gray matter concentration attributable to participation in an MBSR program. Anatomical magnetic resonance (MR) images from 16 healthy, meditation-naïve participants were obtained before and after they underwent the 8-week program. Changes in gray matter concentration were investigated using voxel-based morphometry, and compared with a waiting list control group of 17 individuals. Analyses in a priori regions of interest confirmed increases in gray matter concentration within the left hippocampus. Whole brain analyses identified increases in the posterior cingulate cortex, the temporo-parietal junction, and the cerebellum in the MBSR group compared with the controls. The results suggest that participation in MBSR is associated with changes in gray matter concentration in brain regions involved in learning and memory processes, emotion regulation, self-referential processing, and perspective taking.


Evidence-informed recommendations to reduce dissemination bias in clinical research: conclusions from the OPEN (Overcome failure to Publish nEgative fiNdings) project based on an international consensus meeting.
Dissemination bias in clinical research severely impedes informed decision-making not only for healthcare professionals and patients, but also for funders, research ethics committees, regulatory bodies and other stakeholder groups that make health-related decisions. Decisions based on incomplete and biased evidence cannot only harm people, but may also have huge financial implications by wasting resources on ineffective or harmful diagnostic and therapeutic measures, and unnecessary research. Owing to involvement of multiple stakeholders, it remains easy for any single group to assign responsibility for resolving the problem to others. To develop evidence-informed general and targeted recommendations addressing the various stakeholders involved in knowledge generation and dissemination to help overcome the problem of dissemination bias on the basis of previously collated evidence. Based on findings from systematic reviews, document analyses and surveys, we developed general and targeted draft recommendations. During a 2-day workshop in summer 2013, these draft recommendations were discussed with external experts and key stakeholders, and refined following a rigorous and transparent methodological approach. Four general, overarching recommendations applicable to all or most stakeholder groups were formulated, addressing (1) awareness raising, (2) implementation of targeted recommendations, (3) trial registration and results posting, and (4) systematic approaches to evidence synthesis. These general recommendations are complemented and specified by 47 targeted recommendations tailored towards funding agencies, pharmaceutical and device companies, research institutions, researchers (systematic reviewers and trialists), research ethics committees, trial registries, journal editors and publishers, regulatory agencies, benefit (health technology) assessment institutions and legislators. Despite various recent examples of dissemination bias and several initiatives to reduce it, the problem of dissemination bias has not been resolved. Tailored recommendations based on a comprehensive approach will hopefully help increase transparency in biomedical research by overcoming the failure to disseminate negative findings.


Two-protein signature of novel serological markers apolipoprotein-A2 and serum amyloid alpha predicts prognosis in patients with metastatic renal cell cancer and improves the currently used prognostic survival models.
In metastatic renal cell cancer (mRCC), the Memorial Sloan-Kettering Cancer Center (MSKCC) risk model is widely used for clinical trial design and patient management. To improve prognostication, we applied proteomics to identify novel serological proteins associated with overall survival (OS). Sera from 114 mRCC patients were screened by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). Identified proteins were related to OS. Three proteins were subsequently validated with enzyme-linked immunosorbent assays and immunoturbidimetry. Prognostic models were statistically bootstrapped to correct for overestimation. SELDI-TOF MS detected 10 proteins associated with OS. Of these, apolipoprotein A2 (ApoA2), serum amyloid alpha (SAA) and transthyretin were validated for their association with OS (P = 5.5 x 10(-9), P = 1.1 x 10(-7) and P = 0.0004, respectively). Combining ApoA2 and SAA yielded a prognostic two-protein signature [Akaike's Information Criteria (AIC) = 732, P = 5.2 x 10(-7)]. Including previously identified prognostic factors, multivariable Cox regression analysis revealed ApoA2, SAA, lactate dehydrogenase, performance status and number of metastasis sites as independent factors for survival. Using these five factors, categorization of patients into three risk groups generated a novel protein-based model predicting patient prognosis (AIC = 713, P = 4.3 x 10(-11)) more robustly than the MSKCC model (AIC = 729, P = 1.3 x 10(-7)). Applying this protein-based model instead of the MSKCC model would have changed the risk group in 38% of the patients. Proteomics and subsequent validation yielded two novel prognostic markers and survival models which improved prediction of OS in mRCC patients over commonly used risk models. Implementation of these models has the potential to improve current risk stratification, although prospective validation will still be necessary.


Improved stool DNA integrity method for early colorectal cancer diagnosis.
DNA integrity analysis could represent an alternative approach to the early detection of colorectal cancer. Previously, fluorescence long DNA (FL-DNA) in stools was extracted using a manual approach and analyzed by capillary electrophoresis assay (CE FL-DNA). We aimed to improve diagnostic accuracy using a simpler and more standardized method [Real Time PCR FL-DNA (RT FL-DNA)] for the detection of early malignant lesions in a population undergoing colorectal cancer screening. From 241 stool samples, DNA was extracted using manual and semiautomatic extraction systems and analyzed using FL-DNA tests by CE and RT assays. The RT FL-DNA approach showed slightly higher sensitivity and specificity compared with the CE FL-DNA method. Furthermore, we compared the RT FL-DNA approach with the iFOBT report. Nonparametric ranking statistics were used to analyze the relationship between the median values of RT FL-DNA and the clinicohistopathologic characteristics. The median values of both variables were significantly higher in patients with cancer than in patients with noncancerous lesions. According to the Fagan nomogram results, the iFOBT and FL-DNA methods provided more accurate diagnostic information and were able to identify subgroups at varying risks of cancer. The combination of the semiautomatic extraction system and RT FL-DNA analysis improved the quality of DNA extracted from stool samples. RT FL-DNA shows great potential for colorectal cancer diagnosis as it is a reliable and relatively easy analysis to perform on routinely processed stool samples in combination with iFOBT.


Determination of end point of primary drying in freeze-drying process control.
Freeze-drying is a relatively expensive process requiring long processing time, and hence one of the key objectives during freeze-drying process development is to minimize the primary drying time, which is the longest of the three steps in freeze-drying. However, increasing the shelf temperature into secondary drying before all of the ice is removed from the product will likely cause collapse or eutectic melt. Thus, from product quality as well as process economics standpoint, it is very critical to detect the end of primary drying. Experiments were conducted with 5% mannitol and 5% sucrose as model systems. The apparent end point of primary drying was determined by comparative pressure measurement (i.e., Pirani vs. MKS Baratron), dew point, Lyotrack (gas plasma spectroscopy), water concentration from tunable diode laser absorption spectroscopy, condenser pressure, pressure rise test (manometric temperature measurement or variations of this method), and product thermocouples. Vials were pulled out from the drying chamber using a sample thief during late primary and early secondary drying to determine percent residual moisture either gravimetrically or by Karl Fischer, and the cake structure was determined visually for melt-back, collapse, and retention of cake structure at the apparent end point of primary drying (i.e., onset, midpoint, and offset). By far, the Pirani is the best choice of the methods tested for evaluation of the end point of primary drying. Also, it is a batch technique, which is cheap, steam sterilizable, and easy to install without requiring any modification to the existing dryer.


Electroacupuncture Alleviates Postoperative Cognitive Dysfunction in Aged Rats by Inhibiting Hippocampal Neuroinflammation Activated via Microglia/TLRs Pathway.
Neuroinflammation has been suggested to be involved in the pathogenesis of postoperative cognitive dysfunction (POCD). Electroacupuncture (EA) is an irreplaceable method in traditional Chinese medicine that is used for treating neurodegenerative diseases in clinical and experimental studies. The aim of this study was to examine whether EA improves cognitive dysfunction caused by surgery and to investigate the pathological mechanism of TLR2 and TLR4 in the hippocampus of aged rats. A rat model of POCD was established and treated with EA or minocycline. Both EA- and minocycline-treated rats performed significantly better than untreated operated rats in spatial memory tasks of the Morris water maze (MWM) test, spending comparatively greater amounts of time in the target zone during the probe test. Additionally, decreased levels of proinflammatory cytokines (IL-1β, IL-6, TNF-α, and HMGB1) and decreased TLR2 and TLR4 protein expression in the hippocampus of EA- and minocycline-treated rats were detected. Our data suggested that EA treatment alleviated the cognition performance deficit and neuroinflammation in aged rats following surgery, which may be mediated by inhibiting the expression of hippocampal neuroinflammatory cytokines through the microglia/TLR2/4 pathway.


Laxative effect of probiotic chocolate on loperamide-induced constipation in rats.
The fecal morphology, defecation frequency, bowel function, intestinal motility, and fecal bacterial composition were evaluated to investigate the laxative effect of probiotic chocolate containing Streptococcus thermophilus MG510 and Lactobacillus plantarum LRCC5193 (LYC) on loperamide-induced constipated rats. Daily oral administration of LYC in constipated rats for two weeks was shown to significantly increase (n = 14) the defecation frequency, fecal moisture content, and relative abundance of fecal Lactobacillus and Faecalibacterium prausnitzii. Moreover, histological analysis of the distal colon of constipated rats revealed that LYC treatment can also increase the thickness of the colonic mucosa and muscle layers, and crypt of Lieberkühn. LYC also significantly increased (n = 5) the intestinal motility and modulated (n = 9) mRNA expression levels of colonic ZO-1 and Cldn-1 in the constipated rats. Altogether, these results demonstrate that probiotic chocolate has potential as a dietary adjunct for the treatment of constipation.


Results From the German Fungal Keratitis Registry: Significant Differences Between Cases With and Without a History of Contact Lens Use.
Fungal keratitis (FK) is a serious ophthalmic disease with a potentially devastating outcome that seems to be increasing in recent years. The use of contact lenses (CLs) was evaluated as a risk factor for FK to determine possible differences in course and outcome. Data from 173 cases reported in the German FK registry until August 2019 were evaluated regarding CL behavior, other ophthalmological and general risk factors, age, sex, identified pathogens, conservative and surgical therapy, visual acuity, and findings at admission and follow-up. One hundred seventy-four eyes from 173 patients between 2000 and 2019 were included [64.4% women, median age 54 (39; 72) years]; 49.7% wore CL, of which 81.3% were soft CL, and 50.3% had no history of contact lens use (NCL). CL users were significantly more often women and otherwise healthy (CL: 80.2% vs. NCL: 48.9%; P < 0.0001). The spectrum of pathogens among CL users showed a significantly higher proportion of infections with filamentous pathogens, in particular Fusarium sp. (total filament: CL 69.8% vs. NCL 27.3%; P = 0.0001; Fusarium sp.: CL 50.0% vs. NCL 14.8%; P < 0.0001). Overall, 54.6% required keratoplasty and 8.6% enucleation. CLS are the most important risk factor for FK in Germany. With CLs, typically, the infection is caused by molds, and patients are comparably younger and otherwise healthy. Often, extensive surgery is needed. To evaluate changes in the pathogen and resistance spectrum and to further monitor possible CL-related risk factors, a consistent collection of data remains paramount.


Progressive multifocal leukoencephalopathy in a patient with B-cell lymphoma during rituximab-containing chemotherapy: case report and review of the literature.
Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by the JC polyomavirus. We describe a rare case of PML in a 48-year-old female patient with diffuse large B-cell lymphoma who received rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP) therapy. While she was undergoing five cycles of R-CHOP, she noticed gradually progressive neurological symptoms, such as slurred speech and gait disturbance, and she eventually developed high-grade fever. She also developed Pneumocystis jiroveci pneumonia. The neurological symptoms deteriorated thereafter, and she developed spastic quadriparesis and bulbar palsy. Magnetic resonance imaging showed hyperintensity within the right cerebellar hemisphere on T2-weighted images. Polymerase chain reaction-based tests of the cerebrospinal fluid revealed the presence of the JC virus. Despite intravenous and intrathecal cytarabine treatment, the patient died of PML 5 months after it was diagnosed. Retrospective analysis of her laboratory data showed that her CD4(+) T-cell count before R-CHOP therapy had decreased to 68 microL(-1). Thus, when administering rituximab-containing chemotherapy, even to patients with no prior history of opportunistic infections, attention should be paid to the potential occurrence of PML, particularly in patients with low CD4(+) T-cell counts.


Biological Control Agents Against Fusarium Wilt of Banana.
In the last century, the banana crop and industry experienced dramatic losses due to an epidemic of Fusarium wilt of banana (FWB), caused by Fusarium oxysporum f.sp. cubense (Foc) race 1. An even more dramatic menace is now feared due to the spread of Foc tropical race 4. Plant genetic resistance is generally considered as the most plausible strategy for controlling effectively such a devastating disease, as occurred for the first round of FWB epidemic. Nevertheless, with at least 182 articles published since 1970, biological control represents a large body of knowledge on FWB. Remarkably, many studies deal with biological control agents (BCAs) that reached the field-testing stage and even refer to high effectiveness. Some selected BCAs have been repeatedly assayed in independent trials, suggesting their promising value. Overall under field conditions, FWB has been controlled up to 79% by using Pseudomonas spp. strains, and up to 70% by several endophytes and Trichoderma spp. strains. Lower biocontrol efficacy (42-55%) has been obtained with arbuscular mycorrhizal fungi, Bacillus spp., and non-pathogenic Fusarium strains. Studies on Streptomyces spp. have been mostly limited to in vitro conditions so far, with very few pot-experiments, and none conducted in the field. The BCAs have been applied with diverse procedures (e.g., spore suspension, organic amendments, bioformulations, etc.) and at different stages of plant development (i.e., in vitro, nursery, at transplanting, post-transplanting), but there has been no evidence for a protocol better than another. Nonetheless, new bioformulation technologies (e.g., nanotechnology, formulation of microbial consortia and/or their metabolites, etc.) and tailor-made consortia of microbial strains should be encouraged. In conclusion, the literature offers many examples of promising BCAs, suggesting that biocontrol can greatly contribute to limit the damage caused by FWB. More efforts should be done to further validate the currently available outcomes, to deepen the knowledge on the most valuable BCAs, and to improve their efficacy by setting up effective formulations, application protocols, and integrated strategies.


In Vivo Evidence for a Lactate Gradient from Astrocytes to Neurons.
Investigating lactate dynamics in brain tissue is challenging, partly because in vivo data at cellular resolution are not available. We monitored lactate in cortical astrocytes and neurons of mice using the genetically encoded FRET sensor Laconic in combination with two-photon microscopy. An intravenous lactate injection rapidly increased the Laconic signal in both astrocytes and neurons, demonstrating high lactate permeability across tissue. The signal increase was significantly smaller in astrocytes, pointing to higher basal lactate levels in these cells, confirmed by a one-point calibration protocol. Trans-acceleration of the monocarboxylate transporter with pyruvate was able to reduce intracellular lactate in astrocytes but not in neurons. Collectively, these data provide in vivo evidence for a lactate gradient from astrocytes to neurons. This gradient is a prerequisite for a carrier-mediated lactate flux from astrocytes to neurons and thus supports the astrocyte-neuron lactate shuttle model, in which astrocyte-derived lactate acts as an energy substrate for neurons.


Fluoride therapy and parathyroid hormone activity in osteoporosis.
1. To determine the relationships between parathyroid hormone activity and long-term sodium fluoride therapy in osteoporosis, cytochemical bioassays (for biologically active parathyroid hormone) were performed in 22 osteoporotic control patients and in 18 patients after 15 +/- 10 months of treatment (60 mg of sodium fluoride daily). Ten patients were studied longitudinally by repeated metabolic balances and were therefore common to both groups. All patients were receiving mineral supplements. 2. Cross-sectional data showed a fourfold mean increase in biologically active parathyroid hormone on fluoride treatment (P less than 0.005) together with a 51% increase in serum alkaline phosphatase (P less than 0.005). Longitudinal data showed, in addition, a significant increase in the calcium balance of 2.4 +/- 1.2 (SEM) mmol daily (P less than 0.05) and the development of a positive phosphorus balance (P less than 0.02). 3. Fluoride-treated patients were then analysed in two groups according to the level of biologically active parathyroid hormone. Thirty-two per cent of values were above the upper limit of normal (18 pg/ml). The mean serum alkaline phosphatase level in this group showed no elevation above that of the control patients, the overall rise being accounted for entirely by patients with normal levels of biologically active parathyroid hormone. High levels of biologically active parathyroid hormone were also associated with relative hypophosphataemia (P less than 0.01), relative hypercalciuria (P less than 0.05) and an increased urine/faecal calcium ratio (P less than 0.025). 4. Results show that long-term fluoride and calcium therapy increase biologically active parathyroid hormone in osteoporosis and that excessive parathyroid hormone activity may account for certain features of the refractory state.