Evaluation of chloroquine therapy for vivax and falciparum malaria in southern Sumatra, western Indonesia.
Chloroquine was used as first-line treatment for Plasmodium falciparum or Plasmodium vivax in Indonesia before the initial launch of artemisinin combination therapy in 2004. A study to evaluate efficacies of chloroquine against P. falciparum and P. vivax was undertaken at Lampung in southern Sumatra, western Indonesia in 2002. Patients infected by P. falciparum or P. vivax were treated with 25 mg/kg chloroquine base in three daily doses over 48 hr. Finger prick blood was collected on Days 0, 2, 3, 7, 14, 21 and 28 after starting drug administration. Whole blood chloroquine and its desethyl metabolite were measured on Days-0, -3 and -28, or on the day of recurrent parasitaemia. 42 patients infected by P. falciparum were enrolled, and 38 fullfilled criteria for per protocol analysis. Only six of 38 (16%) showed a response consistent with senstivity to chloroquine. 25 of 32 failures were confirmed resistant by demonstrating chloroquine levels on day of recurrence exceeding the minimally effective concentration (200 ng/mL whole blood). The 28-day cumulative incidence of resistance in P. falciparum was 68% (95% CI: 0.5260 - 0.8306). Thirty one patients infected by P. vivax were enrolled, and 23 were evaluable for per protocol analysis. 15 out of 23 (65%) subjects had persistent or recurrent parasitaemia. Measurement of chloroquine levels confirmed all treatment failures prior to Day-15 as resistant. Beyond Day-15, 4 of 7 recurrences also had drug levels above 100 ng/mL and were classified as resistant. The 28-day cumulative incidence of chloroquine resistance in P. vivax was 43% (95% CI: 0.2715 - 0.6384). These findings confirm persistantly high levels of resistance to chloroquine by P. falciparum in southern Sumatra, and suggest that high-grade and frequent resistance to chloroquine by P. vivax may be spreading westward in the Indonesia archipelago.


The regulation of forkhead/HNF-3beta expression in the Ciona embryo.
The Ciona forkhead/HNF-3beta gene (Ci-fkh) is expressed in the primary axial tissues of the developing tadpole, including the notochord, endoderm, and rudimentary floor plate of the CNS. In an effort to determine the basis for this complex pattern of expression we have conducted a detailed analysis of the Ci-fkh 5'-regulatory region. Different 5' sequences were attached to a lacZ reporter gene and analyzed in electroporated Ciona embryos. A short regulatory sequence (AS) located approximately 1.7 kb upstream of the transcribed region is shown to be essential for expression in all three axial tissues. The proximal 20 bp of the AS contains overlapping Snail repressor elements and a T-box motif. Deleting these sequences causes the loss of reporter gene expression in the endoderm, as well as expanded expression in the neural tube. These results suggest that a T-box gene such as Ci-VegTR activates Ci-fkh expression in the endoderm, while the Ci-Sna repressor excludes expression from the lateral ependymal cells and restricts the Ci-fkh pattern to the rudimentary floor plate in ventral regions of the neural tube. We also present evidence for Ci-fkh positive autofeedback, whereby the Ci-Fkh protein binds to critical activator sites within the Ci-fkh 5'-regulatory region and helps maintain high levels of expression. We discuss these results with respect to forkhead/HNF-3beta regulation in vertebrates.


DNA hypomethylation perturbs the function and survival of CNS neurons in postnatal animals.
DNA methyltransferase I (Dnmt1), the maintenance enzyme for DNA cytosine methylation, is expressed at high levels in the CNS during embryogenesis and after birth. Because embryos deficient for Dnmt1 die at gastrulation, the role of Dnmt1 in the development and function of the nervous system could not be studied by using this mutation. We therefore used the cre/loxP system to produce conditional mutants that lack Dnmt1 in neuroblasts of embryonic day 12 embryos or in postmitotic neurons of the postnatal animal. Conditional deletion of the Dnmt1 gene resulted in rapid depletion of Dnmt1 proteins, indicating that the enzyme in postmitotic neurons turns over quickly. Dnmt1 deficiency in postmitotic neurons neither affected levels of global DNA methylation nor influenced cell survival during postnatal life. In contrast, Dnmt1 deficiency in mitotic CNS precursor cells resulted in DNA hypomethylation in daughter cells. Whereas mutant embryos carrying 95% hypomethylated cells in the brain died immediately after birth because of respiratory distress, mosaic animals with 30% hypomethylated CNS cells were viable into adulthood. However, these mutant cells were eliminated quickly from the brain within 3 weeks of postnatal life. Thus, hypomethylated CNS neurons were impaired functionally and were selected against at postnatal stages.


Attachment/detachment hysteresis of fiber-based magnetic grabbers.
We developed an experimental protocol to analyze the behaviour of a model fiber-based magnetic grabber. A fiber is vertically suspended and fixed to the substrate by its upper end. A magnetic droplet is attached to the free end of the fiber and when a permanent magnet approaches the droplet, the fiber is forced to bow and finally jumps to the magnet. It appears that one can flex the micro-fibers by very small micro or even nano-Newton forces. Using this setup, we discovered a hysteresis of fiber attachment/detachment: the pathway of the fiber jumping to and off the magnet depends on the distance between the magnet and the clamped end. This phenomenon was successfully explained by the Euler-Benoulli model of an elastic beam. The observed hysteresis of fiber attachment/detachment was attributed to the multiple equilibrium configurations of the fiber tip placed in a dipole-type magnetic field.


Fatty acid profiles during gametogenesis in sea urchin (Paracentrotus lividus): effects of dietary inputs on gonad, egg and embryo profiles.
The effects of dietary fatty acids on the composition of Paracentrotus lividus gonads were investigated to determine whether dietary inputs affect their relative abundance during gametogenesis. Egg and embryo FA compositions were compared with that of mature gonads to understand how maternal FA is transferred to the offspring. Urchins were fed an experimental Pellet diet in comparison to brown Kelp (Laminaria digitata). FA profiles of diets, gonads, eggs and embryos revealed the presence in gonads of FA that was absent in the diets and/or higher in contents of some long-chain polyunsaturated fatty acid (LC-PUFA). Moreover, some unusual FA, such as non-methylene interrupted (NMI), was found in gonads, eggs and embryos, but not in the diets, suggesting that P. lividus may be capable of synthesizing this FA and accumulating them in the eggs. A description of gonad FA profiles during gametogenesis is reported for the first time and data suggest that eicosapentaenoic and docosahexaenoic acids are accumulated during gametogenesis, while arachidonic acid is highly regulated and is the only LC-PUFA clearly accumulated into the eggs along with NMI. Further studies are required to determine if maternal provisioning of FA has the potential to influence sea urchin production outputs and to increase hatchery profitability.


Chiral evolution of carbon dots and the tuning of laccase activity.
Chirality has attracted extensive attention in many fields ranging from chemistry to life sciences. Carbon dots (CDs) with good biocompatibility and unique photochemical properties have become a new star in the nanocarbon family. Endowed with chirality, CDs will exhibit more marvellous properties and bridge the fields of material chemistry and life sciences tightly. Herein, we report a facile one-step alkali-assisted electrochemical method to fabricate chiral CDs from cysteine (cys). We showed the chiral evolution of CDs with highly symmetrical circular dichroism (CD) signals in the range from 205 to 350 nm. These chiral CDs have been further demonstrated to be capable of tuning the activity of laccase: the l-CDs can improve the activity of the enzyme up to 20.2%, whereas the d-CDs decrease the activity to 10.4%. A series of experiments confirm that it is the synergistic effect of nanosize and chirality of CDs that induces the change in the structure of laccase and thus leads to the tuning of the laccase activity.


Functional interactions between colony-stimulating factors and the insulin family hormones for human myeloid leukemic cells.
We investigated functional interactions between granulocyte-monocyte-colony-stimulating factor (GM-CSF) and the insulin family hormones using the GM-CSF- and insulin-dependent human acute myeloid leukemia cell line AML-193. Recombinant human GM-CSF and insulin enhanced AML-193 cell proliferation 3- and 5-fold, respectively, and showed a synergistic 10-fold increase when added in combination. Insulin-like growth factors I and II (IGFI and IGFII) increased AML-193 cell proliferation 4-fold and 2-fold, respectively, and also demonstrated synergy when combined with GM-CSF. Blocking experiments with monoclonal antibodies against the insulin and IGFI receptors indicated that the proliferative effects of insulin and IGFI were mediated through both their homologous and heterologous receptors. Pertussis toxin and cholera toxin, which ADP ribosylate GTP-binding proteins (G proteins), and the cyclic AMP analogue, dibutyryl cyclic AMP, decreased the proliferation induced by GM-CSF or insulin. Specific receptor binding of 125I-insulin, -IGFI, and -GM-CSF to AML-193 cells was demonstrated and not affected by preincubation with pertussis toxin or cholera toxin. Radiolabeled GM-CSF, insulin, and IGFI did not cross-compete with the heterologous ligands for receptor binding. These studies demonstrate (a) association between receptor binding and proliferative effects of GM-CSF and the insulin family hormones, (b) involvement of the G proteins in signal transduction provoked by these hormones which occurs at a postreceptor-binding level, and (c) synergistic mitogenic interactions between GM-CSF and the insulin family hormones, suggesting that their receptors are linked to divergent signaling mechanisms in addition to sharing G protein-coupled pathways.


Hormone Receptors and Human Epidermal Growth Factor (HER2) Expression in Fine-Needle Aspirates from Metastatic Breast Carcinoma - Role in Patient Management.
Estrogen receptors (ER), progesterone receptors (PR), and epidermal growth factor (HER2) are prognostic and predictive factors for breast carcinoma. We determined them by immunohistochemistry (IHC) on cell blocks from fine-needle aspirates (FNA) of metastatic breast carcinoma to axillary lymphnodes and compared them with that reported in the primary breast carcinoma (PBC) to document any change in their expression for future management. ER, PR, and HER2 by IHC and HER2 oncogene by fluorescent in-situ hybridization (FISH) were studied on cell blocks of FNA of axillary lymphnodes in 53 of 94 PBC cases from 2012 to 2016. In 25 of 38 (65.8%) ER, PR negative PBC the metastasis on FNA was ER, PR+, whereas the 15 (28.3%) ER, PRPBC remained negative. In 10 of 11 (91%) of HER2-IHC+, PBC the metastatic tumor was HER2-IHC+. 7 of 32 (21.9%) HER2-IHC negative PBC were HER2-IHC+ in metastatic tumor. HER2-FISH was performed in 37 cases on FNA. Six of 37 were HER2 amplified/positive, whereas 9 and 19 remained equivocal and negative for HER2 copy number, and 3 were not interpretable. All the 6 HER2-FISH+ cases were positive by IHC. In our study, 34.2% of ER, PR+ cases of PBC became ER, PR- in the metastatic tumor and 21.9% of HER2-IHC negative PBC became HER2-IHC+ in the metastatic aspirate. ER, PR, and HER2 by IHC in cell blocks of metastatic lymphnodes are reliable. Change in receptor (34.2%) and HER2 status (21.9%) was documented, which is of clinical significance as these patients warrant a change of management.


Infrared Plasmonic Biosensor for Real-Time and Label-Free Monitoring of Lipid Membranes.
In this work, we present an infrared plasmonic biosensor for chemical-specific detection and monitoring of biomimetic lipid membranes in a label-free and real-time fashion. Lipid membranes constitute the primary biological interface mediating cell signaling and interaction with drugs and pathogens. By exploiting the plasmonic field enhancement in the vicinity of engineered and surface-modified nanoantennas, the proposed biosensor is able to capture the vibrational fingerprints of lipid molecules and monitor in real time the formation kinetics of planar biomimetic membranes in aqueous environments. Furthermore, we show that this plasmonic biosensor features high-field enhancement extending over tens of nanometers away from the surface, matching the size of typical bioassays while preserving high sensitivity.


A family's perspective on living with a highly malignant brain tumor.
The purpose of the study is to describe what it like to live with a highly malignant brain tumor from a family perspective. It is a qualitative study in which 3 families, 3 patients, and 5 next of kin have described their experiences in 15 interviews. The study is prospective, with interviews occurring 2-3 weeks after surgery and 3 and 6 months after the onset of the illness. Inductive content analysis has been employed. The results indicate that when a highly malignant brain tumor is diagnosed, the effect on the family is devastating and there is a state of crisis. Characteristically, there is distancing and a sense of helplessness. The members of the family live from day to day in a state of constant anxiety and fear of losing the patient. The affliction limits the patient's capacity regarding activities of daily life, which increases the burden of the next of kin. The next of kin attempt to cope with their grief by occupying themselves with practical tasks and activities that they believe are meaningful. The family members have only good words to say about their encounter with healthcare staff and about the information given. Negative information that the family have not asked for can cause a long period of frustration and anxiety, and they believe that their hope has been taken away from them.


The rationale and design of the Surgical Treatment for Ischemic Heart Failure (STICH) trial.
The rationale and design of the Surgical Treatment for Ischemic Heart Failure trial is described. Before the Surgical Treatment for Ischemic Heart Failure trial, less than 1000 patients with ischemic cardiomyopathy had been studied in randomized comparisons of medical therapy versus coronary artery bypass grafting. Trial data reflect how these therapies were delivered more than 20 years ago and do not indicate the relative benefits of medical therapy versus coronary artery bypass grafting in contemporary practice. Randomization of consenting patients with heart failure, left ventricular ejection fraction of 0.35 or less, and coronary artery disease is based on whether patients are judged by attending physicians to be candidates only for coronary artery bypass grafting or can be treated with medical therapy without coronary artery bypass grafting. Patients eligible for surgical ventricular reconstruction because of significant anterior wall akinesis or dyskinesis but ineligible for medical therapy are randomly assigned to coronary artery bypass grafting with or without surgical ventricular reconstruction. Patients eligible for medical therapy are randomly assigned between medical therapy only and medical therapy with coronary artery bypass grafting. Patients eligible for all 3 are randomly assigned evenly to medical therapy only, medical therapy and coronary artery bypass grafting, or medical therapy and coronary artery bypass grafting and surgical ventricular reconstruction. Major substudies will examine quality of life, cost-effectiveness, changes in left ventricular volumes, effect of myocardial viability, selected biomarkers, and selected polymorphisms on treatment differences. Enrollment is now complete in both STICH hypotheses. Follow-up will continue until sufficient end points are available to address both hypotheses with at least 90% power. The primary outcome of hypothesis 2 is expected to be reported in 2009. The primary outcome of hypothesis 1 is expected to be reported in 2011. The Surgical Treatment for Ischemic Heart Failure trial is a National Heart, Lung, and Blood Institute-funded multicenter international randomized trial addressing 2 specific primary hypotheses: (1) coronary artery bypass grafting with intensive medical therapy improves long-term survival compared with survival with medical therapy alone, and (2) in patients with anterior left ventricular dysfunction, surgical ventricular reconstruction to a more normal left ventricular size plus coronary artery bypass grafting improves survival free of subsequent hospitalization for cardiac cause when compared with that with coronary artery bypass grafting alone.


Neurocognitive and somatic components of temperature increases during g-tummo meditation: legend and reality.
Stories of g-tummo meditators mysteriously able to dry wet sheets wrapped around their naked bodies during a frigid Himalayan ceremony have intrigued scholars and laypersons alike for a century. Study 1 was conducted in remote monasteries of eastern Tibet with expert meditators performing g-tummo practices while their axillary temperature and electroencephalographic (EEG) activity were measured. Study 2 was conducted with Western participants (a non-meditator control group) instructed to use the somatic component of the g-tummo practice (vase breathing) without utilization of meditative visualization. Reliable increases in axillary temperature from normal to slight or moderate fever zone (up to 38.3°C) were observed among meditators only during the Forceful Breath type of g-tummo meditation accompanied by increases in alpha, beta, and gamma power. The magnitude of the temperature increases significantly correlated with the increases in alpha power during Forceful Breath meditation. The findings indicate that there are two factors affecting temperature increase. The first is the somatic component which causes thermogenesis, while the second is the neurocognitive component (meditative visualization) that aids in sustaining temperature increases for longer periods. Without meditative visualization, both meditators and non-meditators were capable of using the Forceful Breath vase breathing only for a limited time, resulting in limited temperature increases in the range of normal body temperature. Overall, the results suggest that specific aspects of the g-tummo technique might help non-meditators learn how to regulate their body temperature, which has implications for improving health and regulating cognitive performance.


External quality assessment schemes for glucose measurements in Germany: factors for successful participation, analytical performance and medical impact.
Determination of blood glucose concentration is one of the most important measurements in clinical chemistry worldwide. Analyzers in central laboratories (CL) and point-of-care tests (POCT) are both frequently used. In Germany, regular participation in external quality assessment (EQA) schemes is mandatory for laboratories performing glucose testing. Glucose testing data from the two German EQAs "Reference Institute for Bioanalytics" (RfB) and "INSTAND - Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien" (Instand) were analyzed from 2012 to 2016. Multivariable odds ratios (OR) for the probability to reach a "good" result were calculated. Imprecision and bias were determined and clinical risk of measurement errors estimated. The device employed was the most important variable required for a "good" performance in all EQAs. Additional participation in an EQA for CL automated analyzers improved performance in POCT EQAs. The reciprocal effect was less pronounced. New participants performed worse than experienced participants especially in CL EQAs. Imprecision was generally smaller for CL, but some POCT devices reached a comparable performance. Large lot-to-lot differences occurred in over 10% of analyzed cases. We propose the "bias budget" as a new metric to express the maximum allowable bias that still carries acceptable medical risk. Bias budgets were smallest and clinical risks of errors greatest in the low range of measurement 60-115 mg/dL (3.3-6.4 mmol/L) for most devices. EQAs help to maintain high analytical performances. They generate important data that serve as the foundation for learning and improvement in the laboratory healthcare system.


Transport and adsorption under liquid flow: the role of pore geometry.
We study here the interplay between transport and adsorption in porous systems with complex geometries under fluid flow. Using a lattice Boltzmann scheme extended to take into account the adsorption at solid/fluid interfaces, we investigate the influence of pore geometry and internal surface roughness on the efficiency of fluid flow and the adsorption of molecular species inside the pore space. We show how the occurrence of roughness on pore walls acts effectively as a modification of the solid/fluid boundary conditions, introducing slippage at the interface. We then compare three common pore geometries, namely honeycomb pores, inverse opal, and materials produced by spinodal decomposition. Finally, we quantify the influence of those three geometries on fluid transport and tracer adsorption. This opens perspectives for the optimization of materials' geometries for applications in dynamic adsorption under fluid flow.


Outcomes of Hip Arthroscopy With Concomitant Periacetabular Osteotomy, Minimum 5-Year Follow-Up.
To report minimum 5-year follow-up results of concomitant hip arthroscopy followed by periacetabular osteotomy (PAO) to treat acetabular dysplasia and intra-articular pathology, such as femoroacetabular impingement syndrome and labral tears. Data were prospectively collected from October 2010 to December 2012. Patients were included in this study if they underwent concomitant hip arthroscopy and PAO and if they had preoperative scores documented for the following measures: modified Harris Hip Score (mHHS), Nonarthritic Hip Score (NAHS), Hip Outcome Score-Sports-Specific Subscale (HOS-SSS), and pain on a visual analog scale (VAS). Patients who underwent reverse PAO to address acetabular retroversion were excluded. Follow-up was considered complete with these outcomes collected after surgery, as well as the abbreviated International Hip Outcome Tool and patient satisfaction on a 0-10 scale. Significance was set at P = .05. Sixteen patients were eligible, all of whom had complete follow-up at a minimum of 5 years after surgery. There were 13 female subjects. The average age of the patients was 23.5 ± 6.8 years (range, 12.3-35.3 years), and the average body mass index was 24.3 ± 5.6 (range, 14.8-34.2). The mean lateral center-edge angle increased from 14.2° to 31.8° (P < .0001), and the anterior center-edge angle increased from 11.9° to 28.6° (P < .0001). The Tönnis angle of acetabular inclination decreased from 19.3° to 2.6° (P < .0001). The alpha angle decreased from 55.7° to 41.0° (P < .0001). All preoperative radiographs were Tönnis ≤1, and there was no progression of arthritis in radiographs taken at the latest clinical visit. All patient-reported outcomes scores demonstrated significant improvement from preoperative baseline to the minimum 5-year follow-up scores (mHHS, P < .001; NAHS, P < .001; HOS-SSS, P = .001). The VAS score decreased from a preoperative mean of 5.8 to 3.1 at the latest follow-up (P = .007). No conversion to total hip arthroplasty was reported. Concomitant hip arthroscopy and PAO appears to be a safe and effective procedure with favorable mid-term outcomes that are durable compared to the short-term. Level IV, case series.


Direct neurophysiological evidence for a role of the human anterior cingulate cortex in central command.
The role of the anterior cingulate cortex (ACC) is still controversial. The ACC has been implicated in such diverse functions as cognition, arousal and emotion in addition to motor and autonomic control. Therefore the ACC is the ideal candidate to orchestrate cardiovascular performance in anticipation of perceived skeletal activity. The aim of this experiment was to investigate whether the ACC forms part of the neural network of central command whereby cardiovascular performance is governed by a top-down mechanism. Direct local field potential (LFP) recordings were made using intraparenchymal electrodes in six human ACC's to measure changes in neuronal activity during performance of a motor task in which anticipation of exercise was uncoupled from skeletal activity itself. Parallel cardiovascular arousal was indexed by electrocardiographic changes in heart rate. During anticipation of exercise, ACC LFP power within the 25-60 Hz frequency band increased significantly by 21% compared to rest (from 62.7 μV2/Hz (±SE 4.94) to 76.0μV2/Hz (±SE 7.24); p = 0.004). This 25-60 Hz activity increase correlated with a simultaneous heart rate increase during anticipation (Pearson's r = 0.417, p = 0.016). We provide the first invasive electrophysiological evidence to support the role of the ACC in both motor preparation and the top-down control of cardiovascular function in exercise. This further implicates the ACC in the body's response to the outside world and its possible involvement in such extreme responses as emotional syncope and hyperventilation. In addition we describe the frequency at which the neuronal ACC populations perform these tasks in the human.


Can AI Help Reduce Disparities in General Medical and Mental Health Care?
As machine learning becomes increasingly common in health care applications, concerns have been raised about bias in these systems' data, algorithms, and recommendations. Simply put, as health care improves for some, it might not improve for all. Two case studies are examined using a machine learning algorithm on unstructured clinical and psychiatric notes to predict intensive care unit (ICU) mortality and 30-day psychiatric readmission with respect to race, gender, and insurance payer type as a proxy for socioeconomic status. Clinical note topics and psychiatric note topics were heterogenous with respect to race, gender, and insurance payer type, which reflects known clinical findings. Differences in prediction accuracy and therefore machine bias are shown with respect to gender and insurance type for ICU mortality and with respect to insurance policy for psychiatric 30-day readmission. This analysis can provide a framework for assessing and identifying disparate impacts of artificial intelligence in health care.


Statins: pleiotropic regulators of cardiovascular redox state.
Lipid-lowering treatment with statins is one of the most effective therapeutic strategies in cardiovascular medicine because they reduce cardiovascular risk in both primary and secondary prevention. Despite the well-established links between low-density lipoprotein and cardiovascular risk, the clinical benefit from statin treatment is not fully explained by their lipid-lowering potential. A number of pleiotropic effects of statins have been described over the past decade, and their ability to suppress global oxidative stress is probably one of the most important mechanisms by which they exert their beneficial effects on the cardiovascular system. In this Forum, there are review articles discussing the molecular mechanisms by which statins modify redox signaling in the vasculature and the heart. They exert direct effects on the vascular wall and the myocardium or indirect by targeting the interactions between the cardiovascular system and adipose tissue or circulating cell types. The review articles in this Forum follow a translational approach and link the molecular mechanisms by which statins modify cardiovascular redox signaling with their clinical benefit in the prevention and treatment of cardiovascular diseases.


Probing the Tumor Suppressor Function of BAP1 in CRISPR-Engineered Human Liver Organoids.
The deubiquitinating enzyme BAP1 is a tumor suppressor, among others involved in cholangiocarcinoma. BAP1 has many proposed molecular targets, while its Drosophila homolog is known to deubiquitinate histone H2AK119. We introduce BAP1 loss-of-function by CRISPR/Cas9 in normal human cholangiocyte organoids. We find that BAP1 controls the expression of junctional and cytoskeleton components by regulating chromatin accessibility. Consequently, we observe loss of multiple epithelial characteristics while motility increases. Importantly, restoring the catalytic activity of BAP1 in the nucleus rescues these cellular and molecular changes. We engineer human liver organoids to combine four common cholangiocarcinoma mutations (TP53, PTEN, SMAD4, and NF1). In this genetic background, BAP1 loss results in acquisition of malignant features upon xenotransplantation. Thus, control of epithelial identity through the regulation of chromatin accessibility appears to be a key aspect of BAP1's tumor suppressor function. Organoid technology combined with CRISPR/Cas9 provides an experimental platform for mechanistic studies of cancer gene function in a human context.


Ionic effects on the proton transfer mechanism in aqueous solutions.
Proton dissociation (PD) reactions of weak acids and proton transfer (PT) processes in aqueous solutions are strongly influenced by ions. However, a detailed molecular picture that describes how ions affect the rates of PD and PT processes is still missing. Here, we utilize time-resolved fluorescence spectroscopy combined with quantum chemical calculations to investigate the excited-state proton transfer (ESPT) reaction of a photoacid in aqueous metal chloride solutions. The activation energy (Ea) for the ESPT of the photoacid increases with increasing charge density of cations (ρcat). The local hydrogen bond (H-bond) structure of the photoacid in the ionic hydration shell is strongly related to both the Ea and the ρcat. Most importantly, the proton's positive charge in the transition state, which is delocalized through the H-bonded water channel, is more destabilized with an increase in the ρcat, leading to a higher Ea. Our experimental and computational results allow us to elucidate the underlying mechanism for the ionic effect on PD and the subsequent PT process at the molecular level.


Clinical Reasoning Assessment Methods: A Scoping Review and Practical Guidance.
An evidence-based approach to assessment is critical for ensuring the development of clinical reasoning (CR) competence. The wide array of CR assessment methods creates challenges for selecting assessments fit for the purpose; thus, a synthesis of the current evidence is needed to guide practice. A scoping review was performed to explore the existing menu of CR assessments. Multiple databases were searched from their inception to 2016 following PRISMA guidelines. Articles of all study design types were included if they studied a CR assessment method. The articles were sorted by assessment methods and reviewed by pairs of authors. Extracted data were used to construct descriptive appendixes, summarizing each method, including common stimuli, response formats, scoring, typical uses, validity considerations, feasibility issues, advantages, and disadvantages. A total of 377 articles were included in the final synthesis. The articles broadly fell into three categories: non-workplace-based assessments (e.g., multiple-choice questions, extended matching questions, key feature examinations, script concordance tests); assessments in simulated clinical environments (objective structured clinical examinations and technology-enhanced simulation); and workplace-based assessments (e.g., direct observations, global assessments, oral case presentations, written notes). Validity considerations, feasibility issues, advantages, and disadvantages differed by method. There are numerous assessment methods that align with different components of the complex construct of CR. Ensuring competency requires the development of programs of assessment that address all components of CR. Such programs are ideally constructed of complementary assessment methods to account for each method's validity and feasibility issues, advantages, and disadvantages.


Management of chronic heart failure: perceived needs of general practitioners in light of the new general medical services contract.
Despite the existence of several chronic heart failure (CHF) guidelines the treatment of patients with CHF is suboptimal. The new general medical services (GMS) contract in primary care has only three specific performance indicators for patients with left ventricular dysfunction. The aim of this current questionnaire survey was to assess the views of general practitioners (GPs) on CHF treatments and services in light of the new GMS contract. All local GPs (717) were sent a questionnaire. Fifty three per cent were returned. Forty five per cent of GPs had access to a community CHF nurse. Having read a national guideline (SIGN) and having the support of a CHF nurse did not seem to affect the knowledge of GPs in terms of perceived benefits of drug treatments. GPs with access to a specialist CHF nurse service attached more importance to it than those with no specialist nurse (p = 0.003). Most GPs were aware of the existence of a national guideline but many had not read it. There was little or no difference in the knowledge level for various evidence based treatments between GPs who had or had not read the guideline suggesting that reading guidelines may not be a key factor in determining knowledge. Support for a specialist CHF nurse was higher among GPs who already had this service, suggesting that this service is valued. The new GMS contract may improve identification and diagnosis of patients with CHF but there is a danger that it may fall short of ensuring optimal treatment for patients with CHF.


Maltodextrin modified liposomes for drug delivery through the blood-brain barrier.
Central nervous system acting drugs, when administered intravenously, cannot show their effect in the brain due to the difficulty in crossing the blood-brain barrier (BBB). Levodopa is one of those drugs that are used to treat Parkinson's disease. In this study, a new liposomal levodopa delivery system that is modified with maltodextrin was developed in order to target and enhance transport through the BBB. An antioxidant, glutathione, was co-loaded in liposomes as a supportive agent and its effect on liposome stability and delivery was investigated. Glutathione co-loading had a positive effect on the viabilities of 3T3 and SH-SY5Y cells. Maltodextrin targeted liposomes showed high in vitro levodopa passage in the parallel artificial membrane permeability assay and had superior binding to MDCK cells. Results suggest that maltodextrin modification of liposomes is an effective way of targeting the BBB and the developed liposomal formulation would improve brain delivery of central nervous system agents.


Cross-Cultural Investigation of Male Gait Perception in Relation to Physical Strength and Speed.
Previous research documents that men and women can accurately judge male physical strength from gait, but also that the sexes differ in attractiveness judgments of strong and weak male walkers. Women's (but not men's) attractiveness assessments of strong male walkers are higher than for weak male walkers. Here, we extend this research to assessments of strong and weak male walkers in Chile, Germany, and Russia. Men and women judged videos of virtual characters, animated with the walk movements of motion-captured men, on strength and attractiveness. In two countries (Germany and Russia), these videos were additionally presented at 70% (slower) and 130% (faster) of their original speed. Stronger walkers were judged to be stronger and more attractive than weak walkers, and this effect was independent of country (but not sex). Women tended to provide higher attractiveness judgments to strong walkers, and men tended to provide higher attractiveness judgments to weak walkers. In addition, German and Russian participants rated strong walkers most attractive at slow and fast speed. Thus, across countries men and women can assess male strength from gait, although they tended to differ in attractiveness assessments of strong and weak male walkers. Attractiveness assessments of male gait may be influenced by society-specific emphasis on male physical strength.


Output correction factors for nine small field detectors in 6 MV radiation therapy photon beams: a PENELOPE Monte Carlo study.
To determine detector-specific output correction factors,[Formula: see text], in 6 MV small photon beams for air and liquid ionization chambers, silicon diodes, and diamond detectors from two manufacturers. Field output factors, defined according to the international formalism published byAlfonso et al. [Med. Phys. 35, 5179-5186 (2008)], relate the dosimetry of small photon beams to that of the machine-specific reference field; they include a correction to measured ratios of detector readings, conventionally used as output factors in broad beams. Output correction factors were calculated with the PENELOPE Monte Carlo (MC) system with a statistical uncertainty (type-A) of 0.15% or lower. The geometries of the detectors were coded using blueprints provided by the manufacturers, and phase-space files for field sizes between 0.5 × 0.5 cm(2) and 10 × 10 cm(2) from a Varian Clinac iX 6 MV linac used as sources. The output correction factors were determined scoring the absorbed dose within a detector and to a small water volume in the absence of the detector, both at a depth of 10 cm, for each small field and for the reference beam of 10 × 10 cm(2). The Monte Carlo calculated output correction factors for the liquid ionization chamber and the diamond detector were within about ± 1% of unity even for the smallest field sizes. Corrections were found to be significant for small air ionization chambers due to their cavity dimensions, as expected. The correction factors for silicon diodes varied with the detector type (shielded or unshielded), confirming the findings by other authors; different corrections for the detectors from the two manufacturers were obtained. The differences in the calculated factors for the various detectors were analyzed thoroughly and whenever possible the results were compared to published data, often calculated for different accelerators and using the EGSnrc MC system. The differences were used to estimate a type-B uncertainty for the correction factors. Together with the type-A uncertainty from the Monte Carlo calculations, an estimation of the combined standard uncertainty was made, assigned to the mean correction factors from various estimates. The present work provides a consistent and specific set of data for the output correction factors of a broad set of detectors in a Varian Clinac iX 6 MV accelerator and contributes to improving the understanding of the physics of small photon beams. The correction factors cannot in general be neglected for any detector and, as expected, their magnitude increases with decreasing field size. Due to the reduced number of clinical accelerator types currently available, it is suggested that detector output correction factors be given specifically for linac models and field sizes, rather than for a beam quality specifier that necessarily varies with the accelerator type and field size due to the different electron spot dimensions and photon collimation systems used by each accelerator model.


Bone assessment after total knee arthroplasty by dual-energy X-ray absorptiometry: analysis protocol and reproducibility.
Bone quality is important for the success of joint prostheses implantation, and the assessment of bone density after total knee arthroplasty by means of dual-energy X-ray absorptiometry may be useful for monitoring implant stability. The aim of this study is to suggest a validated analysis protocol for the assessment of bone status after total knee arthroplasty. A dedicated densitometric analysis protocol of five regions of interest was designed, and 10 subjects who had received an uncemented knee prosthesis (8 females and 2 males, aged 55-74 years) underwent three consecutive scans in posteroanterior and lateral projections, with repositioning after each scan to test the suitability and reproducibility of the protocol. The reproducibility of the measurement of bone mineral content and density in the femoral and tibial regions ranged, respectively, from 2.1% to 4.1%, from 0.9% to 2.6% for the posteroanterior scans, and from 2.7% to 5.6% and from 2.3% to 4.7% for the lateral scans, depending on the considered region. Our results confirm that the suggested protocol allows precise assessment of bone mineral content and density, and that dual-energy X-ray absorptiometry is reliable for the evaluation of bone mass around prosthetic implants.


Spin crossover and reversible single-crystal to single-crystal transformation behaviour in two cyanide-bridged mixed-valence {FeFe} clusters.
The reactions of tricyanometallate precursors, (Ph3PMe)[(Tp4-Me)Fe(CN)3·0.5CH3CN] (1) (Tp4-Me = tri(4-methyl-pyrazol-1-yl)borate) and (NBu4)[(MeTp)Fe(CN)3] (MeTp = methyltris(pyrazolyl)borate) with the presence of the tetradentate tpa ligand (tpa = tris(2-pyridylmethyl)amine) and Fe(ClO4)2·6H2O afford two new cyano-bridged mixed-valence {FeFe} molecular squares: [(Tp4-Me)FeIII(CN)3]2[FeII(tpa)]2·2ClO4·H2O (2·H2O) and [(MeTp)FeIII(CN)3]2[FeII(tpa)]2·2ClO4·CH3OH (3·CH3OH). Solvent-exchange compounds of 3·CH3OH, [(MeTp)FeIII(CN)3]2[FeII(tpa)]2·2ClO4·2H2O (3·2H2O), and their solvent-free form (2 and 3) are also obtained, respectively. The spin crossover (SCO) properties of all compounds are confirmed by detailed structural analyses of the coordination environments of the FeII centres and magnetic susceptibility measurements. All compounds exhibit SCO behaviour near room temperature (T1/2 = 320 K for 2·H2O; 302 K for 2; 292 K for 3·CH3OH; 306 K for 3·2H2O and 290 K for 3) and reversible single-crystal to single-crystal (SC-SC) transformations induced by guest desorption and resorption or solvent exchange. The transition temperature close to room temperature can be tuned by the dehydration and re-hydration processes. The structure-property analysis discloses that the distorted {Fe4(μ-CN)4} core and deviations of bent ∠Fe-N[triple bond, length as m-dash]C angles play a key role in tuning the transition temperature of these similar mixed-valence {FeFe} complexes.


Narratives can motivate environmental action: the Whiskey Creek ocean acidification story.
Even when environmental data quantify the risks and benefits of delayed responses to rapid anthropogenic change, institutions rarely respond promptly. We propose that narratives complementing environmental datasets can motivate responsive environmental policy. To explore this idea, we relate a case study in which a narrative of economic loss due to regionally rapid ocean acidification-an anthropogenic change-helped connect knowledge with action. We pose three hypotheses to explain why narratives might be particularly effective in linking science to environmental policy, drawing from the literature of economics, environmental policy, and cognitive psychology. It seems that yet-untold narratives may hold similar potential for strengthening the feedback between environmental data and policy and motivating regional responses to other environmental problems.


A Comparative Evaluation of Accuracy of New-generation Electronic Apex Locator with Conventional Radiography to determine Working Length in Primary Teeth: An in vivo Study.
The purpose of this study was to evaluate the accuracy of a new-generation electronic apex locator (iPex) to determine working length in primary teeth with or without root resorption as compared with the conventional radiographic method. A sample of 30 primary posterior teeth which are indicated for pulpectomy were selected for the study. After obtaining the informed consent from the parents, local anesthesia was administered. Access cavity was prepared with no.10 round bur. Initial exploration of the canals was done with no.10 K-file. Pulp was extirpated with a barbed broach followed by thorough irrigation of the canals with 0.9% saline. Initially, working length was obtained with iPex (new-generation by Nakanishi International) apex locator using no.10 K-file, which was then compared with conventional radiographic method (Ingle's method). A total of 65 canals were available for the measurement. The data were analyzed using Statistical Analysis system and t-tests were carried out. There was no statistically significant difference found when using iPex apex locator for working length determination as compared with that of conventional radiographic method (p = 0.511). Working length determined by iPex apex locator is comparable with that of conventional radiographic method, hence, can be used as an alternative in determining the working length of primary teeth. Bhat KV, Shetty P, Anandakrishna L. A Comparative Evaluation of Accuracy of New-generation Electronic Apex Locator with Conventional Radiography to determine Working Length in Primary Teeth: An in vivo Study. Int J Clin Pediatr Dent 2017;10(1):34-36.


Comparison of Optimised MDI versus Pumps with or without Sensors in Severe Hypoglycaemia (the Hypo COMPaSS trial).
Severe hypoglycaemia (SH) is one of the most feared complications of type 1 diabetes (T1DM) with a reported prevalence of nearly 40%. In randomized trials of Multiple Daily Injections (MDI) and Continuous Subcutaneous Insulin Infusion (CSII) therapy there is a possible benefit of CSII in reducing SH. However few trials have used basal insulin analogues as the basal insulin in the MDI group and individuals with established SH have often been excluded from prospective studies. In published studies investigating the effect of Real Time Continuous Glucose Monitoring (RT-CGM) benefit in terms of reduced SH has not yet been demonstrated. The primary objective of this study is to elucidate whether in people with T1DM complicated by impaired awareness of hypoglycaemia (IAH), rigorous prevention of biochemical hypoglycaemia using optimized existing self-management technology and educational support will restore awareness and reduce risk of recurrent SH. This is a multicentre prospective RCT comparing hypoglycaemia avoidance with optimized MDI and CSII with or without RT-CGM in a 2×2 factorial design in people with type 1 diabetes who have IAH. The primary outcome measure for this study is the difference in IAH (Gold score) at 24 weeks. Secondary outcomes include biomedical measures such as HbA1c, SH incidence, blinded CGM analysis, self monitored blood glucose (SMBG) and response to hypoglycaemia in gold standard clamp studies. Psychosocial measures including well-being and quality of life will also be assessed using several validated and novel measures. Analysis will be on an intention-to-treat basis. Most existing RCTs using this study's interventions have been powered for change in HbA1c rather than IAH or SH. This trial will demonstrate whether IAH can be reversed and SH prevented in people with T1DM in even those at highest risk by using optimized conventional management and existing technology. ISRCTN52164803 Eudract No: 2009-015396-27.


BMI1 is a therapeutic target in recurrent medulloblastoma.
Medulloblastoma (MB) is the most frequent malignant pediatric brain tumor, representing 20% of newly diagnosed childhood central nervous system malignancies. Although advances in multimodal therapy yielded a 5-year survivorship of 80%, MB still accounts for the leading cause of childhood cancer mortality. In this work, we describe the epigenetic regulator BMI1 as a novel therapeutic target for the treatment of recurrent human Group 3 MB, a childhood brain tumor for which there is virtually no treatment option beyond palliation. Current clinical trials for recurrent MB patients based on genomic profiles of primary, treatment-naive tumors will provide limited clinical benefit since recurrent metastatic MBs are highly genetically divergent from their primary tumor. Using a small molecule inhibitor against BMI1, PTC-028, we were able to demonstrate complete ablation of self-renewal of MB stem cells in vitro. When administered to mice xenografted with patient tumors, we observed significant reduction in tumor burden in both local and metastatic compartments and subsequent increased survival, without neurotoxicity. Strikingly, serial in vivo re-transplantation assays demonstrated a marked reduction in tumor initiation ability of recurrent MB cells upon re-transplantation of PTC-028-treated cells into secondary recipient mouse brains. As Group 3 MB is often metastatic and uniformly fatal at recurrence, with no current or planned trials of targeted therapy, an efficacious targeted agent would be rapidly transitioned to clinical trials.


Species interactions and the structure of complex communication networks.
A universal challenge faced by animal species is the need to communicate effectively against a backdrop of heterospecific signals. It is often assumed that this need results in signal divergence to minimize interference among community members, yet previous support for this idea is mixed, and few studies have tested the opposing hypothesis that interactions among competing species promote widespread convergence in signaling regimes. Using a null model approach to analyze acoustic signaling in 307 species of Amazonian birds, we show that closely related lineages signal together in time and space and that acoustic signals given in temporal or spatial proximity are more similar in design than expected by chance. These results challenge the view that multispecies choruses are structured by temporal, spatial, or acoustic partitioning and instead suggest that social communication between competing species can fundamentally organize signaling assemblages, leading to the opposite pattern of clustering in signals and signaling behavior.


Garlic and Heart Disease.
Thousands of studies have been published based on animal and human studies evaluating garlic's effects and safety. We reviewed the available literature investigating the effects of garlic supplements on hypertension, hypercholesterolemia, C-reactive protein (CRP), pulse wave velocity (PWV), and coronary artery calcium (CAC), as well as available data on side effects. We searched PubMed for all human studies using medical subject heading words through 30 May 2013 and assessed relevant review articles and original studies. Only double-blind, randomized, controlled trials and meta-analyses of double-blind, randomized, controlled trials were included. The review of articles and data extraction were performed by 2 independent authors, with any disagreements resolved by consensus. Garlic supplementation reduced blood pressure by 7-16 mm Hg (systolic) and 5-9 mm Hg (diastolic) (4 meta-analyses and 2 original studies). It reduced total cholesterol by 7.4-29.8 mg/dL (8 meta-analyses). The most consistent benefits were shown in studies that used aged garlic extract (AGE). A few small studies that used AGE also showed favorable effects on CAC, CRP, and PWV. Although garlic is generally safe, rare adverse reactions have been documented with limited causality established. We conclude that garlic supplementation has the potential for cardiovascular protection based on risk factor reduction (hypertension and total cholesterol) and surrogate markers (CRP, PWV, and CAC) of atherosclerosis. Larger studies are warranted to evaluate these effects further.


The number of alleles in DNA mixtures with related contributors.
The maximum allele count (MAC) across loci and the total allele count (TAC) are often used to gauge the number of contributors to a DNA mixture. Computational strategies that predict the total number of alleles in a mixture arising from a certain number of contributors of a given population have been developed. Previous work considered the restricted case where all of the contributors to a mixture are unrelated. We relax this assumption and allow mixture contributors to be related according to a pedigree. We introduce an efficient computational strategy. This strategy based on first determining a probability distribution on the number of independent alleles per locus, and then conditioning on this distribution to compute a distribution of the number of distinct alleles per locus. The distribution of the number of independent alleles per locus is obtained by leveraging the Identical by Descent (IBD) pattern distribution which can be computed from the pedigree. We explain how allelic dropout and a subpopulation correction can be accounted for in the calculations.


Drop-to-drop variation in the cellular components of fingerprick blood: implications for point-of-care diagnostic development.
Blood obtained via fingerprick is commonly used in point-of-care assays, but few studies have assessed variability in parameters obtained from successive drops of fingerprick blood, which may cause problems for clinical decision making and for assessing accuracy of point-of-care tests. We used a hematology analyzer to analyze the hemoglobin concentration, total WBC count, three-part WBC differential, and platelet count in six successive drops of blood collected from one fingerprick from each of 11 donors, and we used a hemoglobinometer to measure the hemoglobin concentration of 10 drops of fingerprick blood from each of 7 donors. The average percent coefficient of variation (CV) for successive drops of fingerprick blood was higher by up to 3.4 times for hemoglobin, 5.7 times for WBC count, 3 times for lymphocyte count, 7.7 times for granulocyte count, and 4 times for platelets than in venous controls measured using a hematology analyzer. The average percent CV for fingerprick blood was up to 5 times higher for hemoglobin than venous blood measured using a point-of-care hemoglobinometer. Fluctuations in blood parameters with increasing volume of fingerprick blood are within instrument variability for volumes equal to or greater than 60 to 100 μL. These data suggest caution when using measurements from a single drop of fingerprick blood.


Prioritising the application of genomic medicine.
The clinical translation of genomic sequencing is hampered by the limited information available to guide investment into those areas where genomics is well placed to deliver improved health and economic outcomes. To date, genomic medicine has achieved its greatest successes through applications to diseases that have a high genotype-phenotype correlation and high penetrance, with a near certainty that the individual will develop the condition in the presence of the genotype. It has been anticipated that genomics will play an important role in promoting population health by targeting at-risk individuals and reducing the incidence of highly prevalent, costly, complex diseases, with potential applications across screening, prevention, and treatment decisions. However, where primary or secondary prevention requires behavioural changes, there is currently very little evidence to support reduction in disease incidence. A better understanding of the relationship between genomic variation and complex diseases will be necessary before effective genomic risk identification and management of the risk of complex diseases in healthy individuals can be carried out in clinical practice. Our recommended approach is that priority for genomic testing should focus on diseases where there is strong genotype-phenotype correlation, high or certain penetrance, the effects of the disease are serious and near-term, there is the potential for prevention and/or treatment, and the net costs incurred are acceptable for the health gains achieved.


Novel hormone treatment of benign metastasizing leiomyoma: an analysis of five cases and literature review.
To evaluate novel hormonal therapies in patients with unresectable benign metastasizing leiomyoma (BML) disease. Case series. National Institutes of Health (NIH). Five subjects with the diagnosis of BML based on imaging and/or histopathologic diagnosis. Four patients were treated with single or combination therapy of leuprolide acetate and/or an aromatase inhibitor. One patient was treated with an antiprogestin (CDB-2914). Response to therapy was measured by tumor burden on cross-sectional imaging employing RECIST (Response Evaluation Criteria in Solid Tumors) 1.1 guidelines. Four patients treated with single or combination therapy of leuprolide acetate and/or an aromatase inhibitor demonstrated stable disease with reduction in tumor burden. The fifth patient treated with antiprogestin (CDB-2914) had degeneration of her tumor, progression of its size, and an improvement in symptoms. Hormone treatment with GnRH agonist and/or aromatase inhibition may be a therapeutic option to reduce tumor burden in unresectable BML disease or for those patients who wish to avoid surgical intervention. RECIST 1.1 guidelines, while traditionally used to evaluate tumor response to cancer therapeutics, may be useful in evaluating BML tumor burden response to hormone therapy.


Therapy with immunosuppressive drugs and biological agents and use of contraception in patients with rheumatic disease.
To investigate the attitude of patients towards immunosuppressive and biological drugs in relation to reproduction and the outcome of pregnancies exposed to these drugs. We performed 2 postal surveys in regard to immunosuppressive drugs and reproduction, one in patients with rheumatic disease, the second in Swiss rheumatologists. Among the 237 female patients and the 189 male patients contacted for the survey, 72% of women and 40% of men returned the questionnaire. Ninety-four women and 47 men had received one or several immunosuppressive or biological agents during the years 2000-2005. Correct advice in regard to drugs and necessary birth control had been given to 84% of women. Advice to men was more inconsistent. One-third of women and 50% of men treated with potentially teratogenic drugs methotrexate (MTX) or leflunomide had not practiced birth control. The surveys of rheumatologists and patients disclosed 66 pregnancies under therapy with immunosuppressive and biological drugs with successful outcomes in 73%. However, 20% of pregnancies in women occurred under treatment with MTX and leflunomide. Issues regarding drugs and reproduction are not always sufficiently discussed with female and male patients. The increasing use of combination therapies containing MTX necessitates ensuring that advice regarding birth control is followed in order to avoid pregnancies exposed to potentially fetotoxic drugs.


Automated immunostaining of cell smears: an alternative to flow cytometry.
To assess the applicability of an automated tissue immunostainer machine for the phenotypic analysis of peripheral blood and bone marrow aspirate smears. Air-dried smears of peripheral blood and bone marrow from normal individuals and 14 patients with haematological malignancies were stained, following fixation, with a range of antibodies to haemopoietic antigens using both immunoperoxidase and immuno-alkaline phosphatase methods on the Bond-maX (Leica Microsystems) automated immunostaining machine. Automated protocols used for staining formalin-fixed paraffin-embedded tissue could be applied to cell smears, giving high quality staining with excellent cytological detail and antigenic preservation for an extensive antibody panel. Optimal quality (morphological preservation and antigen detection without background staining) was obtained with an alkaline phosphatase method and Fast Red chromogenic substrate. Both cell surface and intracellular (cytoplasmic and nuclear) antigens could be detected and gave the expected reactivity. Automated immunostaining is possible for haematological smears. It generates consistent high quality staining, with the exception of surface immunoglobulin, and is applicable to haematological and, potentially, cytological smears. It provides a practical alternative to flow cytometry and will have particular application when smears are available and there is no suitable sample for flow cytometry.


Molecular targets of the natural antioxidant pterostilbene: effect on protein kinase C, caspase-3 and apoptosis in human neutrophils in vitro.
Pterostilbene, a naturally occurring phenolic derivative, exhibits various pharmacological effects, e.g. anti-cancerous, antioxidant, anti-inflammatory and anti-diabetic. Based on our previous study, we assessed the cellular and molecular effects of pterostilbene on human neutrophils and in cell free systems. Experimental and theoretical molecular descriptors of stilbene derivatives were also determined. We assessed the antioxidant properties of pterostilbene using cell free system and computational methods. The effect of pterostilbene on protein kinase C activation/phosphorylation was detected by special anti-phospho protein kinase C antibodies. Membrane associated changes determining the life span of neutrophils and human recombinant caspase-3 assay were examined. Pterostilbene possessed comparable antioxidant properties as resveratrol in cell free system. Computational methods were used to establish the molecular characteristics of stilbene derivatives. The values of electronic parameters suggest a slight enhancement of electron donor properties of pterostilbene compared to resveratrol. Phosphorylation and thus activation of protein kinase C alpha/beta II in activated neutrophils was not decreased by pterostilbene. Pterostilbene in concentrations of 10-100 μM was found to inhibit the activity of human caspase-3 purified enzyme and did not influence cell viability significantly. Pterostilbene, an analog of resveratrol, was identified as a good natural antioxidant compound. However, reducing the oxidative burst of human neutrophils during their activation in vitro with pterostilbene does not include protein kinase C phosphorylation pathway. Pterostilbene showed dose dependent activation/inhibition of caspase-3 enzyme activity.


Kinetics of cytokine mRNA production in the brains of mice with progressive toxoplasmic encephalitis.
C57BL/10 ScSn (B10) mice infected orally with Toxoplasma gondii were killed on days 5, 10, 15, 20 and 30 post infection and their brains excised. These were either used to count total tissue cyst numbers or divided for RNA purification and histopathological studies. The first signs of inflammation were on day 10 post infection, before the appearance of cysts in the brain, and correlating with the appearance of activated astrocytes. These mice had a mild meningitis with areas of encephalitis. Small numbers of cyst stages were first observed in the brain on day 15 and by day 20 the cyst numbers had increased dramatically but were not always associated with inflammation. After this time point, total cyst numbers did not increase significantly though there developed a marked variation in tissue cyst size with larger cysts becoming more numerous. The use of the polymerase chain reaction to assist in the amplification of brain RNA allowed the characterization of the kinetics of cytokine production within the brains of these animals. Only IL-1 alpha was found to be expressed constitutively in control mice. Transcripts for other cytokines associated with activated monocytes, microglial cells and astrocytes [tumor necrosis factor (TNF)-alpha and interleukin (IL)-6] were present on day 10, (IL-6) and day 15 (TNF-alpha) post infection. Thereafter, these cytokines were present in all infected animals. Of the T cell-associated cytokines, IL-4, a characteristic product of the T helper (Th)2 cell subset, was detected on days 10 and 15, while granulocyte macrophage colony stimulating factor (GM-CSF) which can be produced not only by this cell type but also by Th1 cells and CD8+ T cells, was also present on day 15 but not thereafter. Transcripts for interferon (IFN)-gamma, present from day 15 post infection, were probably produced by CD8+ T cells, as IL-2 which would indicate Th1 cell involvement was only detected 30 days after infection. The continual presence of IFN-gamma and TNF-alpha, cytokines with reported anti-toxoplasmic activity, in the CNS of B10 mice throughout the latter half of the experimental period did not diminish the severity of infection. These results indicate that the CD4+ Th2 subset may allow a rapid rise in cyst numbers and so be important in determining susceptibility to toxoplasmic encephalitis.


Isolated sphenoid sinus disease: differential diagnosis and management.
To review the current literature on isolated disease of sphenoid sinus, and to describe the various approaches to the sphenoid sinus. Due to its posterior location and subtle presenting symptoms, sphenoid lesions, in the past, have often been missed and were reported as rare occurrences. However, with the availability of current diagnostic modalities such as endoscopy and imaging techniques such as computed tomography (CT) and magnetic resonance imaging (MRI), diseases of the sphenoid sinus are now more frequently diagnosed. Since the introduction of endoscopic techniques in the mid 1980s, various endoscopic approaches to the sphenoid sinus, depending on the extent and type of disease, have been described. These approaches were subsequently 'extended' to remove pituitary tumours, and certain diseases of the parasellar region and the petrous apex. More recently, further extension of these approaches has led to the development of endoscopic skull base surgery, using the sphenoid sinus as the 'gateway' to the anterior, middle and the posterior cranial fossa. These approaches require a clear understanding of the anatomy of the sphenoid sinus and its surrounding regions. This paper presents a review of the various diseases of the sphenoid sinus and describes the approaches to the sinus with special emphasis on the transpterygoid approach to the lateral recess of the sphenoid sinus.


Trypanosoma spp. in Swedish game animals.
Serum and blood samples from 36 game animals, shot during the hunting seasons 2007-2009, were collected and analyzed for the presence of Trypanosoma spp. by three methods: isolation, polymerase chain reaction (PCR), and serology. Only fissiped animals were included, four different ruminants and wild boar. Trypanosomes could be isolated from two of the animals, and eight had detectable parasite DNA. Seven animals had high titers of anti-trypanosoma IgG antibodies. The two isolated strains, one from roe dear and one from European elk, were determined to Trypanosoma theileri by partial DNA sequencing of the 18S ribosomal gene. In the seven boars, no Trypanosoma were detected, but four out of seven strongly positive serological samples came from this group. This is the first study in Scandinavia on the presence of Trypanosoma in game animals. The results indicate that trypanosomiasis is frequently occurring among Swedish game animals.


Quantitative assessment of angiogenesis in murine antigen-induced arthritis by intravital fluorescence microscopy.
Inhibition of angiogenesis might be a therapeutic approach to prevent joint destruction caused by the overgrowing synovial tissue during chronic joint inflammation. The aim of this study was to investigate angiogenesis in the knee joint of mice with antigen-induced arthritis (AIA) by means of intravital microscopy. In 14 mice (C57BL6/129Sv) intravital microscopic assessment was performed on day 8 after AIA induction in two groups (controls, AIA). Synovial tissue was investigated by intravital fluorescence microscopy using FITC-dextran (150 kD). Quantitative assessment of vessel density was performed according to the following categories: functional capillary density (FCD, vessels <10 microm in diameter), functional vessel density (FVD, vessels >10 microm) and FVD of vessels with angiogenic criteria (convoluted vessels, abrupt changes of diameter, vessels which are generated by sprouting and progressively pruned and remodelled). Microvessel count was performed using immunohistochemistry. There was no significant difference in FCD between the control group (337 +/- 9 cm/cm2; mean +/- SEM) and the AIA group (359 +/- 13 cm/cm2). The density of vessels larger than 10 microm diameter was significantly increased in animals with AIA (135 +/- 10 vs. 61 +/- 5 cm/cm2 in control). The density of blood vessels with angiogenic criteria was enhanced in arthritic animals (79 +/- 17 vs. 12 +/- 2 cm/cm2 in control). There was a significant increase in the microvessel count in arthritic animals (297 +/- 25 vs. 133 +/- 16 mm(-2) in control). These findings demonstrate that angiogenesis in murine AIA can be assessed quantitatively using intravital microscopy. Further studies will address antiangiogenic strategies in AIA.


Estimating Morbidity Rates Based on Routine Electronic Health Records in Primary Care: Observational Study.
Routinely recorded electronic health records (EHRs) from general practitioners (GPs) are increasingly available and provide valuable data for estimating incidence and prevalence rates of diseases in the population. This paper describes how we developed an algorithm to construct episodes of illness based on EHR data to calculate morbidity rates. The goal of the research was to develop a simple and uniform algorithm to construct episodes of illness based on electronic health record data and develop a method to calculate morbidity rates based on these episodes of illness. The algorithm was developed in discussion rounds with two expert groups and tested with data from the Netherlands Institute for Health Services Research Primary Care Database, which consisted of a representative sample of 219 general practices covering a total population of 867,140 listed patients in 2012. All 685 symptoms and diseases in the International Classification of Primary Care version 1 were categorized as acute symptoms and diseases, long-lasting reversible diseases, or chronic diseases. For the nonchronic diseases, a contact-free interval (the period in which it is likely that a patient will visit the GP again if a medical complaint persists) was defined. The constructed episode of illness starts with the date of diagnosis and ends at the time of the last encounter plus half of the duration of the contact-free interval. Chronic diseases were considered irreversible and for these diseases no contact-free interval was needed. An algorithm was developed to construct episodes of illness based on routinely recorded EHR data to estimate morbidity rates. The algorithm constitutes a simple and uniform way of using EHR data and can easily be applied in other registries.


Cyberknife stereotactic radiosurgery for treatment of atypical (WHO grade II) cranial meningiomas.
The optimal management of subtotally resected atypical meningiomas is unknown. To perform a retrospective review of patients with residual or recurrent atypical meningiomas treated with stereotactic radiosurgery (SRS). Twenty-five patients were treated, either immediately after surgery (n = 15) or at the time of radiographic progression or treatment failure (n = 10). SRS was delivered to with a median marginal dose of 22 Gy (range, 16-30) in 1 to 4 fractions (median, 1), targeting a median tumor volume of 5.3 cm³ (range, 0.3-26.0). With a median follow-up time of 28 months (range, 3-67), the 12-, 24-, and 36-month actuarial local and regional control rates for all patients were 94%, 94%, 74%, and 90%, 90%, 62%, respectively. There were 2 cases of radiation toxicity. On univariate analysis, the number of recurrences before SRS (P = .046), late SRS (ie, waiting until tumor progression to initiate treatment) (P = .03), and age at treatment ≥ 60 years (P = .01) were significant predictors of recurrence. Of the 20 radiation-naïve patients, 2 patients failed with the targeted lesion and 3 elsewhere in the resection bed, resulting in 12-, 24- and 36-month actuarial local and regional control rates of 100%, 100%, 73% and 93%, 93%, 75%, respectively. The overall locoregional control rates at 12, 24, and 36 months were 93%, 93%, and 54%, respectively. Irradiation of the entire postoperative tumor bed may not be necessary for the majority of patients with subtotally resected atypical meningiomas. Patients in this series achieved outcomes comparable to that of historical control rates for larger volume, conventionally fractionated radiotherapy.


Estimated risk of HIV acquisition and practice for preventing occupational exposure: a study of healthcare workers at Tumbi and Dodoma Hospitals, Tanzania.
Healthcare workers (HCWs) are at risk of acquiring human immuno-deficiency virus (HIV) and other infections via exposure to infectious patients' blood and body fluids. The main objective of this study was to estimate the risk of HIV transmission and examine the practices for preventing occupational exposures among HCWs at Tumbi and Dodoma Hospitals in Tanzania. This study was carried out in two hospitals, namely, Tumbi in Coast Region and Dodoma in Dodoma Region. In each facility, hospital records of occupational exposure to HIV infection and its management were reviewed. In addition, practices to prevent occupational exposure to HIV infection among HCWs were observed. The estimated risk of HIV transmission due to needle stick injuries was calculated to be 7 cases per 1,000,000 HCWs-years. Over half of the observed hospital departments did not have guidelines for prevention and management of occupational exposure to HIV infections and lacked well displayed health and safety instructions. Approximately, one-fifth of the hospital departments visited failed to adhere to the instructions pertaining to correlation between waste materials and the corresponding colour coded bag/container/safety box. Seventy four percent of the hospital departments observed did not display instructions for handling infectious materials. Inappropriate use of gloves, lack of health and safety instructions, and lack of use of eye protective glasses were more frequently observed at Dodoma Hospital than at Tumbi Hospital. The poor quality of the hospital records at the two hospitals hampered our effort to characterise the risk of HIV infection acquisition by HCWs. Greater data completeness in hospital records is needed to allow the determination of the actual risk of HIV transmission for HCWs. To further reduce the risk of HIV infection due to occupational exposure, hospitals should be equipped with sufficient personal protective equipment (PPE) and HCWs should be reminded of the importance of adhering to universal precautions.


Magneto-induced anisotropy in magnetic colloids of superparamagnetic magnetite nanoparticles in an external magnetic field.
Here we report a study of chain formation and the magnetic anisotropy induced by them in suspensions of slightly anisotropic Fe3O4 nanoparticles in water and in aqueous V2O5 suspensions. An investigation into the magnetization of the fluid and frozen suspensions, the application of dynamic light scattering techniques and the observation of the magnetic anisotropy in the frozen magnetically aligned samples allowed us to confirm the existence of chains of Fe3O4 in both suspensions. Our study shows that the magneto-induced anisotropy appearing in magnetic fields in colloids with Fe3O4 particles is mainly due to many particle (chain) magneto-induced anisotropy, but not due to single particle magneto-induced anisotropy connected with particle shape anisotropy. In other words, the single particle magneto-induced anisotropy is much smaller than the many particle (chain) anisotropy. The anisometry of the chains provides effective coupling with the nonmagnetic V2O5 component of the suspension and results in its strong sensitivity to the magnetic field.


Imaging enzymes at work: metabolic mapping by enzyme histochemistry.
For the understanding of functions of proteins in biological and pathological processes, reporter molecules such as fluorescent proteins have become indispensable tools for visualizing the location of these proteins in intact animals, tissues, and cells. For enzymes, imaging their activity also provides information on their function or functions, which does not necessarily correlate with their location. Metabolic mapping enables imaging of activity of enzymes. The enzyme under study forms a reaction product that is fluorescent or colored by conversion of either a fluorogenic or chromogenic substrate or a fluorescent substrate with different spectral characteristics. Most chromogenic staining methods were developed in the latter half of the twentieth century but still find new applications in modern cell biology and pathology. Fluorescence methods have rapidly evolved during the last decade. This review critically evaluates the methods that are available at present for metabolic mapping in living animals, unfixed cryostat sections of tissues, and living cells, and refers to protocols of the methods of choice.


Submillimeter spectroscopy for chemical analysis with absolute specificity.
A sensor based on rotational signatures in the submillimeter (SMM) region is described. This sensor uses frequency synthesis techniques in the region around 10 GHz, with nonlinear diode frequency multiplication to 210-270 GHz. This provides not only a nearly ideal instrument function, but also frequency control and agility that significantly enhance the performance of the spectrometer as a sensor. The SMM frequencies provide significantly stronger absorptions and broader spectroscopic coverage than lower-frequency microwave systems. Among the characteristics of the sensor are absolute specificity, low atmospheric clutter, good sensitivity, and near-term paths to systems that are both compact and very inexpensive.


Prebiotic evaluation of cocoa-derived flavanols in healthy humans by using a randomized, controlled, double-blind, crossover intervention study.
The absorption of cocoa flavanols in the small intestine is limited, and the majority of the flavanols reach the large intestine where they may be metabolized by resident microbiota. We assessed the prebiotic potential of cocoa flavanols in a randomized, double-blind, crossover, controlled intervention study. Twenty-two healthy human volunteers were randomly assigned to either a high-cocoa flavanol (HCF) group (494 mg cocoa flavanols/d) or a low-cocoa flavanol (LCF) group (23 mg cocoa flavanols/d) for 4 wk. This was followed by a 4-wk washout period before volunteers crossed to the alternant arm. Fecal samples were recovered before and after each intervention, and bacterial numbers were measured by fluorescence in situ hybridization. A number of other biochemical and physiologic markers were measured. Compared with the consumption of the LCF drink, the daily consumption of the HCF drink for 4 wk significantly increased the bifidobacterial (P < 0.01) and lactobacilli (P < 0.001) populations but significantly decreased clostridia counts (P < 0.001). These microbial changes were paralleled by significant reductions in plasma triacylglycerol (P < 0.05) and C-reactive protein (P < 0.05) concentrations. Furthermore, changes in C-reactive protein concentrations were linked to changes in lactobacilli counts (P < 0.05, R(2) = -0.33 for the model). These in vivo changes were closely paralleled by cocoa flavanol-induced bacterial changes in mixed-batch culture experiments. This study shows, for the first time to our knowledge, that consumption of cocoa flavanols can significantly affect the growth of select gut microflora in humans, which suggests the potential prebiotic benefits associated with the dietary inclusion of flavanol-rich foods. This trial was registered at clinicaltrials.gov as NCT01091922.


Phytotracker, an information management system for easy recording and tracking of plants, seeds and plasmids.
A large number of different plant lines are produced and maintained in a typical plant research laboratory, both as seed stocks and in active growth. These collections need careful and consistent management to track and maintain them properly, and this is a particularly pressing issue in laboratories undertaking research involving genetic manipulation due to regulatory requirements. Researchers and PIs need to access these data and collections, and therefore an easy-to-use plant-oriented laboratory information management system that implements, maintains and displays the information in a simple and visual format would be of great help in both the daily work in the lab and in ensuring regulatory compliance. Here, we introduce 'Phytotracker', a laboratory management system designed specifically to organise and track plasmids, seeds and growing plants that can be used in mixed platform environments. Phytotracker is designed with simplicity of user operation and ease of installation and management as the major factor, whilst providing tracking tools that cover the full range of activities in molecular genetics labs. It utilises the cross-platform Filemaker relational database, which allows it to be run as a stand-alone or as a server-based networked solution available across all workstations in a lab that can be internet accessible if desired. It can also be readily modified or customised further. Phytotracker provides cataloguing and search functions for plasmids, seed batches, seed stocks and plants growing in pots or trays, and allows tracking of each plant from seed sowing, through harvest to the new seed batch and can print appropriate labels at each stage. The system enters seed information as it is transferred from the previous harvest data, and allows both selfing and hybridization (crossing) to be defined and tracked. Transgenic lines can be linked to their plasmid DNA source. This ease of use and flexibility helps users to reduce their time needed to organise their plants, seeds and plasmids and to maintain laboratory continuity involving multiple workers. We have developed and used Phytotracker for over five years and have found it has been an intuitive, powerful and flexible research tool in organising our plasmid, seed and plant collections requiring minimal maintenance and training for users. It has been developed in an Arabidopsis molecular genetics environment, but can be readily adapted for almost any plant laboratory research.


Modulation of memory formation by stimulus content: specific role of the medial prefrontal cortex in the successful encoding of social pictures.
It is unclear whether the involvement of the medial prefrontal cortex (mPFC) during encoding is restricted to the evaluative processing of to-be-encoded stimuli or if it is instead actively engaged during memory formation. The difficulty of assessing the contribution of the mPFC to encoding based on previous neuroimaging studies partly arises from the use of several types of stimuli, such as emotional or social ones. These different types of stimulus content could differently modulate mPFC activity during memory formation and thus partly explain the variable contribution of this region to encoding. Using emotional/neutral and social/nonsocial pictures, we conducted an event-related functional magnetic resonance imaging study using a subsequent memory paradigm as the main analytical strategy. We observed that the brain activity in the dorsal and orbital mPFC is significantly and specifically predictive of the successful encoding of social compared with nonsocial pictures. In contrast, the activity in the amygdala specifically predicts the successful encoding of emotional compared with neutral pictures. The modulation of the mPFC by social information in a memory encoding context could be associated with the initiation of self-referential processes whose contribution is to enhance memory formation.


The impact of helical computed tomography on the diagnosis of unsuspected inflammatory bowel disease in the large bowel.
Inflammatory bowel diseases (IBD) are diagnoses that can be easily overlooked clinically. The aim of this study was to investigate if CT is able to make a contribution to the identification of previously unsuspected cases of IBD. We retrospectively identified cases in which the possibility of an IBD was raised in CT reports (over a 4-year period), by utilising a keyword search of the radiology database. Cases with a previously known or suspected IBD were rigorously excluded by review of case notes, and endoscopic, radiological, histological and microbiological findings. The CT images of the identified cases were reviewed by a blinded observer to document the extent of bowel wall thickening, the location of lesion(s), and presence of peri-colic fat abnormality, ascites and/or collections. The observer also attempted to corroborate the presence, and to identify the type, of IBD based on the CT appearances alone. Thirty-five cases (out of approximately 19,000 body CTs performed) of clinically unsuspected IBD were identified, of which 27 underwent further investigation. An IBD was confirmed in 48% (13 of 27): Crohn's disease (n=6), ulcerative colitis (n=2), pseudomembranous colitis (n=3) and other (n=2), of which 70% (9 of 13) were correctly typed by the reviewer. Inflammatory bowel disease was not substantiated in the remainder (14 of 27), although 7 of these had other bowel pathologies: diverticular disease (n=4); and carcinoma (n=3). Prospectively determining the presence, and furthermore type, of IBD on CT is challenging largely because of the considerable overlap in the appearances of the individual IBDs and indeed of normality. Nevertheless, CT is able to identify clinically unsuspected cases and radiologists should be alert to this treatable and not infrequently elusive diagnosis.


Analyzing schizophrenia by DNA microarrays.
To understand the pathological processes of schizophrenia, we must embrace the analysis of the diseased human brain: we will never be able to recapitulate the pathology of uniquely human disorders in an animal model. Based on the outcome of the transcriptome profiling experiments performed to date, it appears that schizophrenia is associated with a global gene expression disturbance across many cortical regions. In addition, transcriptome changes are present in multiple cell types, including specific subclasses of principal neurons, interneurons, and oligodendrocytes. Furthermore, transcripts related to synaptic transmission, energy metabolism, and inhibitory neurotransmission are routinely found underexpressed in the postmortem brain tissue of subjects with schizophrenia. To put these transcriptome data in biological context, we must make our data publicly available and report our findings in a proper, expanded Minimum Information About a Microarray Experiment format. Cell-type specific expression profiling and sequencing-based transcript assessments should be expanded, with particular attention to understanding splice-variant changes in various mental disorders. Deciphering the pathophysiology of mental disorders depends on integrating data from across many research fields and techniques. Leads from postmortem transcriptome profiling will be essential to generate model animals, perform tissue culture experiments, and develop or evaluate novel drugs to treat this devastating disorder.


The sperm factor: paternal impact beyond genes.
The fact that sperm carry more than the paternal DNA has only been discovered just over a decade ago. With this discovery, the idea that the paternal condition may have direct implications for the fitness of the offspring had to be revisited. While this idea is still highly debated, empirical evidence for paternal effects is accumulating. Male condition not only affects male fertility but also offspring early development and performance later in life. Several factors have been identified as possible carriers of non-genetic information, but we still know little about their origin and function and even less about their causation. I consider four possible non-mutually exclusive adaptive and non-adaptive explanations for the existence of paternal effects in an evolutionary context. In addition, I provide a brief overview of the main non-genetic components found in sperm including DNA methylation, chromatin modifications, RNAs and proteins. I discuss their putative functions and present currently available examples for their role in transferring non-genetic information from the father to the offspring. Finally, I identify some of the most important open questions and present possible future research avenues.


Urzymology: experimental access to a key transition in the appearance of enzymes.
Urzymes are catalysts derived from invariant cores of protein superfamilies. Urzymes from both aminoacyl-tRNA synthetase classes possess sophisticated catalytic mechanisms: pre-steady state bursts, significant transition-state stabilization of both amino acid activation, and tRNA acylation. However, they have insufficient specificity to ensure a fully developed genetic code, suggesting that they participated in synthesizing statistical proteins. They represent a robust experimental platform from which to articulate and test hypotheses both about their own ancestors and about how they, in turn, evolved into modern enzymes. They help reshape numerous paradigms from the RNA World hypothesis to protein structure databases and allostery.


Phase I/II study of galiximab, an anti-CD80 antibody, for relapsed or refractory follicular lymphoma.
This multicenter, dose-escalation study evaluates the safety, pharmacokinetics, and efficacy of galiximab (anti-CD80 monoclonal antibody) in patients with relapsed or refractory follicular lymphoma. Patients had follicular lymphoma that had relapsed or failed to respond to primary therapy; the majority (90%) presented with stage III or IV disease. Four weekly intravenous infusions of galiximab were administered at doses of 125, 250, 375, or 500 mg/m2. Thirty-seven patients received galiximab treatment and were evaluated for safety; 35 were assessable for response. Antibody infusions were safe and well tolerated with no dose-limiting toxicities. A total of 22 (60%) of 37 patients experienced adverse events related to galiximab. All but one of the events were grade 1 or 2; the most common were fatigue, nausea, and headache. Cytopenias were rare; only one patient experienced anemia and febrile neutropenia, which were unrelated to galiximab and resolved after treatment. No patient developed antigaliximab antibody formation. The mean serum half-life ranged from 13 to 24 days. The overall response rate was 11% (two complete responses and two partial responses). Time to best response was delayed (months 3, 6, 9, and 12). Twelve patients (34%) maintained stable disease. Nearly half of all patients (49%) had a decrease in indicator lesions. Two responders remain on study without progression (22 and 24.4 months). The favorable safety profile of galiximab and evidence of single-agent biologic activity and dose-dependent pharmacokinetics support further evaluation of galiximab as a treatment for follicular lymphoma, possibly in combination with other lymphoma therapies.


Putative Abscopal Effect in Three Patients Treated by Combined Radiotherapy and Modulated Electrohyperthermia.
Purpose: True abscopal responses from radiation therapy are extremely rare; the combination of immune checkpoint inhibitors with radiation therapy has led to more reports of the abscopal effect, but even in this setting, the genuine magnitude remains unknown and is still considered generally uncommon. We report the occurrence of what appears to be putative, durable abscopal tumor responses with associated auto-immune systemic reactions resulting from the combination of local radiotherapy (RT) and modulated electrohyperthermia (mEHT). Materials and Methods: Data from advanced cancer patients treated palliatively with RT and mEHT between January and December 2017 were collected as part of a post-marketing safety monitoring program of mEHT therapy. We specified a minimum RT dose of 30 Gy and at least four mEHT treatments for reporting toxicities, which was the primary aim of the larger study. Results: Thirty-three patients treated with RT and mEHT, both applied to the same lesion, were included. The median RT dose was 45.5 Gy in 20 fractions (fxs) and the median number of mEHT treatments was 12 (range, 4-20). Most patients had subsequent systemic therapy after one course of RT and mEHT. Three patients (9.1%) developed autoimmune toxicities. Case number 1 received RT and mEHT only; case number 2 had two cycles of concurrent low dose chemotherapy during RT; and case number 3 received concurrent immune checkpoint inhibitors. None of the three patients received any further systemic treatment due to obvious treatment-related autoimmune reactions which occurred rapidly after RT; one had autoimmune hepatitis, one had dermatitis herpetiformis and the third developed severe myasthenia gravis. Interestingly, what we surmise to be long-lasting abscopal responses outside the irradiated area, were noted in all three patients. Conclusion: RT combined with mEHT could putatively result in enhancing immune responsiveness. These preliminary observational findings lead to the generation of a hypothesis that this combination induces both an in-situ, tumor-specific immune reaction and an anti-self-autoimmune reaction, in at least a small proportion of patients, and of those who experience the auto-immune response, tumor response is a concomitant finding. Mechanisms underlying this phenomenon need to be investigated further.


Clinical outcomes of lung transplant recipients with telomerase mutations.
Successful lung transplantation for patients with pulmonary fibrosis from telomerase mutations may be limited by systemic complications of telomerase dysfunction, including myelosuppression, cirrhosis, and malignancy. We describe clinical outcomes in 14 lung transplant recipients with telomerase mutations. Subjects underwent lung transplantation between February 2005 and April 2014 at 5 transplant centers. Data were abstracted from medical records, focusing on outcomes reflecting post-transplant treatment effects likely to be complicated by telomerase mutations. The median age of subjects was 60.5 years (interquartile range = 52.0-62.0), 64.3% were male, and the mean post-transplant observation time was 3.2 years (SD ± 2.9). A mutation in telomerase reverse transcriptase was present in 11 subjects, a telomerase RNA component mutation was present in 2 subjects, and an uncharacterized mutation was present in 1 subject. After lung transplantation, 10 subjects were leukopenic and 5 did not tolerate lymphocyte anti-proliferative agents. Six subjects developed recurrent lower respiratory tract infections, 7 developed acute cellular rejection (A1), and 4 developed chronic lung allograft dysfunction. Eight subjects developed at least 1 episode of acute renal failure and 10 developed chronic renal insufficiency. In addition, 3 subjects developed cancer. No subjects had cirrhosis. At data censorship, 13 subjects were alive. The clinical course for lung transplant recipients with telomerase mutations is complicated by renal disease, leukopenia with intolerance of lymphocyte anti-proliferative agents, and recurrent lower respiratory tract infections. In contrast, cirrhosis was absent, acute cellular rejection was mild, and development of chronic lung allograft dysfunction was comparable to other lung transplant recipients. Although it poses challenges, lung transplantation may be feasible for patients with pulmonary fibrosis from telomerase mutations.


Phase I trial of MK-0752 in children with refractory CNS malignancies: a pediatric brain tumor consortium study.
To estimate the maximum-tolerated dose (MTD), describe dose-limiting toxicities (DLTs), and characterize pharmacokinetic properties of MK-0752, a gamma secretase inhibitor, in children with refractory or recurrent CNS malignancies. MK-0752 was administered once daily for 3 consecutive days of every 7 days at escalating dosages starting at 200 mg/m(2). The modified continual reassessment method was used to estimate the MTD. A course was 28 days in duration. Pharmacokinetic analysis was performed during the first course. Expression of NOTCH and hairy enhancer of split (HES) proteins was assessed in peripheral-blood mononuclear cells (PBMCs) before and following treatment with MK-0752. Twenty-three eligible patients were enrolled: 10 males (median age, 8.1 years; range, 2.6 to 17.7 years) with diagnoses of brainstem glioma (n = 6), ependymoma (n = 8), medulloblastoma/primitive neuroectodermal tumor (n = 4), glioblastoma multiforme (n = 2), atypical teratoid/rhabdoid tumor (n = 1), malignant glioma (n = 1), and choroid plexus carcinoma, (n = 1). Seventeen patients were fully evaluable for toxicity. No DLTs occurred in the three patients enrolled at 200 mg/m(2)/dose. At 260 mg/m(2)/dose, DLTs occurred in two of six patients, both of whom experienced grade 3 ALT and AST. There were no grade 4 toxicities; non-dose-limiting grade 3 toxicities included hypokalemia and lymphopenia. Population pharmacokinetic values (% coefficient of variation) for MK-0752 were apparent oral clearance, 0.444 (38%) L/h/m(2); apparent volume of distribution, 7.36 (24%) L/m(2); and k(a), 0.358 (99%) hr(-1). MK-0752 is well-tolerated in children with recurrent CNS malignancies. The recommended phase II dose using the 3 days on followed by 4 days off schedule is 260 mg/m(2)/dose once daily.


Hydrogenase- and outer membrane c-type cytochrome-facilitated reduction of technetium(VII) by Shewanella oneidensis MR-1.
Pertechnetate, (99)Tc(VII)O(4)(-), is a highly mobile radionuclide contaminant at US Department of Energy sites that can be enzymatically reduced by a range of anaerobic and facultatively anaerobic microorganisms, including Shewanella oneidensis MR-1, to poorly soluble Tc(IV)O(2(s)). In other microorganisms, Tc(VII)O(4)(-) reduction is generally considered to be catalysed by hydrogenase. Here, we provide evidence that although the NiFe hydrogenase of MR-1 was involved in the H(2)-driven reduction of Tc(VII)O(4)(-)[presumably through a direct coupling of H(2) oxidation and Tc(VII) reduction], the deletion of both hydrogenase genes did not completely eliminate the ability of MR-1 to reduce Tc(VII). With lactate as the electron donor, mutants lacking the outer membrane c-type cytochromes MtrC and OmcA or the proteins required for the maturation of c-type cytochromes were defective in reducing Tc(VII) to nanoparticulate TcO(2) x nH(2)O((s)) relative to MR-1 or a NiFe hydrogenase mutant. In addition, reduced MtrC and OmcA were oxidized by Tc(VII)O(4)(-), confirming the capacity for direct electron transfer from these OMCs to TcO(4)(-). c-Type cytochrome-catalysed Tc(VII) reduction could be a potentially important mechanism in environments where organic electron donor concentrations are sufficient to allow this reaction to dominate.